Datasets Archive
Collection
Data produced relating to model train/test cycles which have been archived - due to data versioning or priorities changing
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6 items
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Updated
PMCID
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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KIT proto-oncogene, receptor tyrosine kinase (KIT, CD117) and platelet-derived growth factor-alpha (PDGFRA) are key drivers of gastrointestinal stromal tumors (GIST), but resistance to targeted therapy often arises from tumor protein p53 (p53) alterations and loss of cell cycle control.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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However, the role of p53 status in GIST therapeutic potential has rarely been studied, so this study aimed to employ both wild-type and mutant p53 GIST models to investigate how p53 dysfunction influences the efficacy of p53 pathway-targeted therapies.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The efficacy of the mouse double minute 2 homolog (MDM2) inhibitor (HDM201) and the Wee1 G2 checkpoint kinase (Wee1) inhibitor (adavosertib) was confirmed in both p53 wild-type (p53 WT) and p53 mutant (p53 MT) GIST cells.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The anti-proliferative effects were assessed using the Cell Counting Kit-8 (CCK-8) assay.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Flow cytometry (FACS) and immunoblotting were employed to evaluate apoptosis and the expression of proteins related to drug efficacy.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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These findings were further validated in a xenograft model.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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HDM201 selectively inhibited growth and triggered apoptosis in p53 WT GIST cells, while adavosertib was effective mainly in p53 MT cells.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Western blot analysis revealed that HDM201 increased p53 and p21 levels in p53 WT cells, and adavosertib affected Wee1 and phospho-cdc2 expression in both p53 WT and p53 MT cells.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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In a xenograft mouse model, HDM201 significantly reduced the tumor volume and weight in p53 WT GIST cells, whereas p53 MT tumors showed only a moderate size reduction with adavosertib, without significant changes.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Our results highlight the importance of p53 status in guiding GIST treatment.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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p53 WT tumors respond to MDM2 inhibitors, while p53 MT tumors show greater sensitivity to Wee1 inhibitors, supporting p53 pathway targeting as a promising strategy for GIST patients.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Gastrointestinal stromal tumors (GISTs) belong to a category of mesenchymal tumors that originate in the gastrointestinal tract.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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More than 90% of GISTs overexpress the KIT proto-oncogene, receptor tyrosine kinase (KIT, CD117) protein [1–3].
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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A deeper understanding of GISTs has emerged with the identification of KIT expression and the KIT gene (c-Kit) .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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An additional 5%–10% of GISTs contain mutations that overexpress platelet-derived growth factor-alpha (PDGFRA) .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Small-molecule targeted therapies utilizing tyrosine kinase inhibitors (TKIs) have been developed to treat GIST .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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While the majority of GISTs are characterized by mutations in the KIT or PDGFRA kinase genes, approximately 5%–10% of GISTs deviate from this pattern and lack mutations in either KIT or PDGFRA.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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These tumors were categorized as KIT/PDGFRA wild-type (WT) GISTs.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The tumor protein p53 (TP53) gene, which encodes the p53 tumor suppressor protein, is often referred to as the guardian of the genome.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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It is mutated in most human cancers, with mutation frequencies that vary according to the specific cancer type .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Wild-type p53 (p53 WT) protein plays a crucial role in the cellular response to DNA damage by initiating cell cycle arrest, DNA repair, and apoptosis .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The occurrence and potential prognostic relevance of TP53 mutations (p53 MT) have been explored in a spectrum of cancer types [7–9].
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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However, in gastrointestinal stromal tumors (GISTs), p53 has received less attention, as GIST oncogenesis is primarily driven by the dysregulation of KIT and PDGFRA .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Key molecular events, such as cyclin-dependent kinase inhibitor 2A (CDKN2A) loss, mouse double minute 2 homolog (MDM2) overexpression, and p53 inactivation, are critical in GIST progression .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Given its role in cell cycle regulation and DNA damage response, p53 is implicated in the progression of high-risk GISTs.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Although TP53 mutations are uncommon in GIST, they are more prevalent in high-risk cases and are significantly associated with poorer relapse-free survival .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Furthermore, p53 expression serves as an independent prognostic factor in advanced GISTs treated with imatinib .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Collectively, these findings highlight the critical role of p53 expression and TP53 mutations in GIST progression.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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In p53 WT high-risk GISTs, the p53 signaling pathway is often disrupted due to MDM2 overexpression, which impairs p53’s tumor-suppressive functions, leading to genomic instability, uncontrolled proliferation, and oncogene activation.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Consequently, targeting MDM2 or other components of the p53 pathway represents a promising therapeutic approach for GISTs .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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These insights support the development of p53 pathway-targeted therapies as a novel strategy for GIST treatment.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Previous studies have revealed that inhibiting Wee1 G2 checkpoint kinase (Wee1) promotes autophagic degradation of KIT, suggesting that targeting Wee1 could offer a novel therapeutic strategy for GIST .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Wee1 plays a critical role in regulating the G2/M cell cycle checkpoint, allowing cancer cells with DNA damage to continue through the replication cycle .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Furthermore, Wee1 inhibition with adavosertib (MK1775) disrupts the G2/M checkpoint by preventing phosphorylation of cyclin-dependent kinase-1 (CDK1, cdc2), thereby inducing apoptosis in p53-mutant ovarian and lung cancer cells .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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p53 transcriptionally activates MDM2, which in turn promotes negative autoregulation of p53 through ubiquitination .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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As a result, MDM2 is classified as an oncogene, and its amplification or overexpression has the potential to enhance tumor cell proliferation by suppressing the activity of p53 .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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As MDM2 expression is associated with poor prognosis in GIST, MDM2 inhibitors (nutlin-3) have been shown to suppress growth and induce apoptosis in p53 WT GIST cells .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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HDM201 (siremadlin) is a novel, highly potent, and selective inhibitor of the p53-MDM2 interaction.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The use of MDM2 inhibitors may provide an additional strategy beyond targeted therapy drugs such as imatinib, sunitinib, regorafenib, and ripretinib in future studies .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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This study aimed to confirm the association among p53 status, HDM201, and adavosertib activity in GIST.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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This study used three human GIST cell lines: GIST430 with wild-type p53 (p53 WT), GIST882, and GIST-T1 with mutant p53 (p53 MT).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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All cell lines were kindly provided by Dr. Nai Jung Chiang (Taipei Veterans General Hospital, Taipei, Taiwan).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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GIST430 cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM) (Gibco, 12440053, Waltham, MA, USA) supplemented with 20% heat-inactivated fetal bovine serum (FBS) (Gibco, 10437-028).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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GIST882 cells were cultured in Roswell Park Memorial Institute (RPMI) medium 1640 (Gibco, 11875-085) supplemented with 20% FBS, and GIST-T1 cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, 11965-984) supplemented with 10% FBS.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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All three GIST cell lines were cultured with 100 μg/mL streptomycin and 100 μg/mL penicillin (Gibco, 15140-122) in a humidified atmosphere containing 5% CO2 at 37°C.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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All the cell lines used in this study were verified to be free of mycoplasma contamination and authenticated through short tandem repeat (STR) profiling.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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GIST430, GIST882, and GIST-T1 cells were seeded in 96-well plates (3 × 10/well) and incubated overnight, and then treated with HDM201 (obtained from Novartis under a material transfer agreement (MTA), GST0000026313, Basel, Basel-Stadt, Switzerland) or adavosertib (MedChemExpress, Basel, Basel-Stadt, Switzerland) for 96 h. The gradient concentrations of HDM201 from 0–10 μM and adavosertib from 0–5 μM used in this study were determined based on prior in vitro studies involving cancer models such as GIST, colon cancer, and ovarian cancer .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Cell viability was assessed using the Cell Counting Kit-8 (CCK8, Dojindo Molecular Technologies, Rockville, MD, USA) method, and the optical density was measured at 450 nm using a microplate reader (Synergy HTX Multi-Mode Reader, BioTek, Winooski, VT, USA).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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TP53 siRNA (siTP53-1 sense: 5-CCACCAUCCACUACAACUAdTdT-3, antisense: 5-UAGUUGUA GUGGAUGGUGGdTdT-3 and siTP53-2 sense: 5-GAUGUUCCGAGAGCUGAAUdTdT-3, antisense: 5-AUUCAGCUCUCGGAACAUCdTdT-3) or negative control siRNA (siNC) (sense: 5-UUCU CCGAACGUGUCACGUTT-3, antisense: 5-ACGUGACACGUUCGGAGAATT-3) were transfected into GIST430 cells using DharmaFECT transfection reagent (horizon, Cambridge, UK) for 24 h. Western blotting was performed to confirm knockdown efficacy.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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After successful TP53 knockdown, CCK8, a growth inhibition assay, was performed to evaluate the effect of HDM201 or adavosertib on p53 knockdown GIST430 cells.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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GIST cells, treated with siRNA, HDM201, or adavosertib, were lysed in Pierce RIPA Lysis and Extraction Buffer (Thermo Scientific, Waltham, MA, USA) supplemented with protease inhibitors (Roche, Basel, Switzerland).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The lysates were centrifuged at 12,000× g for 20 min at 4°C, and the protein concentration of the supernatants was measured using the Pierce BCA Protein Assay Kit (Thermo Scientific, 23225).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Total cell lysates were separated on 10% SDS-PAGE gels and transferred to nitrocellulose membranes (Amersham, Cytiva, Marlborough, MA, USA).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween 20 (TBST) at room temperature.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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After blocking, the membranes were incubated overnight at 4°C with primary antibodies, followed by TBST washes and incubation with secondary antibodies, in 5% skim milk for 1–2 h at room temperature.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Protein detection was performed using chemiluminescence with horseradish peroxidase-conjugated goat anti-mouse or anti-rabbit secondary antibodies (1:5000, Jackson ImmunoResearch Laboratories, mouse: #115-035-0031, rabbit: #11-035-003, West Grove, PA, USA), and protein bands were visualized using the UVP ChemStudio PLUS Touch imager (Analytik Jena AG, Jena, Germany).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The primary antibodies used were as follows: p53 (1:1000, GeneTex, GTX34938, Irvine, CA, USA), KIT (1:1000, ABclonal Technology, A0357, Woburn, MA, USA), phospho-KIT (1:1000, ABclonal Technology, AP0385), Wee1 (1:1000, GeneTex, GTX111392), phospho-Wee1 (1:1000, Cell Signaling Technology, #4910, Danvers, MA, USA), cdc2 (1:1000, GeneTex, GTX108120), phospho-cdc2 (1:1000, GeneTex, GTX128155), MDM2 (1:1000, MilliporeSigma, #MABE281, Burlington, MA, USA), p21 (1:1000, Cell Signaling Technology, #2947) and GAPDH (1:20000, GeneTex, GTX627408).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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GIST cells in 6-well plates (4 × 10/well) were treated with 1 μM HDM201 or adavosertib for 48 h. For the cell cycle distribution assay, both floating and adherent cells were collected, fixed with cold 70% ethanol, and incubated with Propidium Iodide (PI)/RNase Staining Solution (Cell Signaling Technology, #4087) for 20 min in the dark at room temperature.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The samples were then analyzed for DNA content using a FACSCalibur flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA) and CellQuest Pro software (Becton Dickinson), with results processed in FlowJo vX software (Becton Dickinson).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The FITC Annexin V Apoptosis Detection Kit I (Becton Dickinson, 556547) was used for the apoptosis assays.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The cells were washed twice with cold 1 × PBS (pH 7.4), resuspended in binding buffer, and stained with fluorescein isothiocyanate (FITC), Annexin V, and PI according to manufacturer’s protocol.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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After 15 min of incubation at room temperature in the dark, the cells were analyzed using a FACSCalibur flow cytometer (Becton Dickinson).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The proportion of early and late apoptotic cells was calculated using FlowJo vX software (Becton Dickinson).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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GIST430, GIST882, and GIST-T1 cells were seeded in white 96-well white plates (1 × 10/well) and treated with 1 μ M HDM201 or adavosertib for 24 h. Caspase-3/7 enzymatic activities were measured using a luminometer (Synergy HTX Multi-Mode Reader, BioTek), after adding a 1:1 ratio of Caspase Glo-3/7 reagent (Promega, G8090, Fitchburg, WI, USA) to growth media and incubating for 60 min.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The animal studies were approved by the Institutional Animal Care and Use Committee (IACUC, No. 2019032003) of Chang Gung Memorial Hospital at Linkou.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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All animal experiments were conducted in accordance with the Guide for the Care and Use of Laboratory Animals .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The animals were housed in an Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC)-approved facility in our hospital, under controlled temperature (24°C) and a 12-h light/dark cycle.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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A total of 20 specific pathogen-free, immunodeficient NonObese Diabetic/Severe Combined Immunodeficiency (NOD/SCID) male mice, aged four weeks and weighing about 20 g, were obtained from BioLASCO Co., Ltd., Taiwan.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Based on previous studies [26–28], 1 × 10 GIST430 or GIST882 cells were suspended in 50 μL of PBS (pH 7.2) mixed with an equal volume of Matrigel (1:1, Corning, 354262, Corning, NY, USA) and then subcutaneously injected into NOD/SCID mice.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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When the average tumor size reached 100 mm, the mice were randomized into two groups (five per group) and treated via oral gavage.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The GIST430 group received either vehicle (0.5% methylcellulose) or HDM201 (100 mg/kg/day, 2 days/week) according to a previous study .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Similarly, the GIST882 group received either vehicle or adavosertib (50 mg/kg/day, 5 days/week) based on a prior study .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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During the treatment period, tumor size, , and pain/distress classifications were monitored twice a week until the largest tumor approached but did not exceed 2 cm in diameter, which was designated as the endpoint.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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On day 11 in our two animal studies, mice were sacrificed for tumor collection.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The results shown in Figs. 1–3 represent data from three independent experiments and were analyzed statistically.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Values are expressed as the mean ± standard error of the mean (SEM).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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All statistical tests were performed using the GraphPad Prism 8 software (GraphPad Software, San Diego, CA, USA).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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p-values represent the results of unpaired t-tests or two-way analysis of variance (ANOVA), and differences with p-values less than 0.05 were considered statistically significant.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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GIST430, GIST882, and GIST-T1 cells were treated with HDM201 (Fig. 1A) and adavosertib (Fig. 1B) for 96 h. Growth inhibition by HDM201 was observed in GIST430 cells, a wild-type p53 cell line, but not in p53 mutated cells, including GIST882 and GIST-T1 cells (Fig. 1A).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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In contrast, adavosertib inhibited the growth of GIST882 and GIST-T1 cells but not of GIST430 cells (Fig. 1B).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Two siRNAs targeting TP53 (siTP53-1 and 2) successfully suppressed the expression of p53 in GIST430 cells (Fig. 1C, the uncropped Western blot images are provided in Fig. S1).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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When GIST430 cells were treated with 1 µM HDM201, the growth inhibitory activity decreased after suppression of wild-type p53 expression indicating that the activity of HDM201 is p53-dependent.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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However, no significant difference was observed in cell survival among the cells treated with adavosertib after p53 knockdown (Fig. 1D).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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The cell proliferation assay showed that HDM201 effectively inhibited the growth of p53 WT cells, whereas adavosertib effectively inhibited the growth of p53 MT cells.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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GIST cells were treated with HDM201 (1 µM) or adavosertib (1 µM) for 48 h. In GIST430 cells, HDM201 decreased the number of S-phase cells, whereas no significant effect was observed in GIST882 and GIST-T1 cells.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Adavosertib treatment led to increased S and G2/M phase arrest in GIST-T1 cells and induced S phase arrest in GIST882 cells (Fig. 2A,B).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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HDM201 significantly increased the sub-G1 population of GIST430 cells but did not affect GIST882 or GIST-T1 cells.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Conversely, adavosertib significantly increased the number of sub-G1 GIST882 and GIST-T1 cells, but not the number of GIST430 cells (Fig. 2A,C).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Flow cytometry staining with annexin V showed that HDM201 treatment enhanced apoptosis in GIST430 cells, and adavosertib promoted early and late apoptotic cell production in GIST882 and GIST-T1 cells (Fig. 3A,B).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Furthermore, caspase 3/7 activity was increased two-fold in GIST430 cells treated with HDM201 (1 µM) compared to the control group, as observed in GIST882 and GIST-T1 cells (Fig. 3C).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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When GIST cells were treated with adavosertib, caspase 3/7 activity tripled in GIST882 and GIST-T1 cells compared to that in both control and GIST430 cells (Fig. 3D).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Cell cycle distribution, annexin V, and caspase 3/7 results showed that HDM201 decreased the S phase population and increased the sub-G1 phase population and apoptosis in p53 WT cells, whereas adavosertib increased apoptosis in p53 MT cells, along with an increase in the sub-G1 phase and induction of S phase and/or G2/M arrest.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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To explore the on-target effects of HDM201 and adavosertib, western blot analysis of the three cell lines treated with these drugs was conducted.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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We analyzed the proteins associated with p53 activity (p53 and p21), which are regulated by MDM2, and cell cycle-related proteins (cdc2, p-cdc2, Wee1, and p-Wee1), which are influenced by adavosertib in these cells (Fig. 4).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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As depicted by western blotting, compared with DMSO-treated GIST cells, all GIST cells treated with adavosertib increased the expression of phosphorylation of Wee1 and decreased the expression of phospho-cdc2.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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In addition, no changes were observed in the expression of Wee1 and cdc2.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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GIST430 treated with HDM201 showed over-expression of p53 and p21; however, no difference was observed in expression between GIST882 and GIST-T1 treated with HDM201 and DMSO.
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Additionally, consistent with previous studies, the use of adavosertib alone did not affect KIT phosphorylation .
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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Wee1 inhibition did not significantly decrease the phosphorylation of KIT (Y721) in Fig. 4 (The uncropped Western blot images are provided in Figs. S2 and S3).
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PMC12573211
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Efficacy of Wee1 G2 Checkpoint Kinase and Mouse Double Minute 2 Homolog Inhibitors in Gastrointestinal Stromal Tumors Determined by p53 Status
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HDM201 reactivated p53 WT but did not influence KIT expression.
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Dataset Card for Dataset Name
Dataset Description
Dataset Summary This dataset has been extracted from Europe PMC (EPMC), a free database offering comprehensive access to life sciences research literature. EPMC aggregates content from various sources, including PubMed, arXiv, and other repositories, and provides open access to millions of scientific articles. This dataset has been generated as part of a project collaboration between Europe PMC, Open Targets, and ChEMBL] at EMBL-EBI.
Dataset Details
The dataset focuses on mentions of cell lines and related entities in biomedical text, such as cell types, related disease mentions, genes proteins etc. This makes it a valuable resource as it can be used for(but not limited to) the following downstream natural language processing (NLP) tasks in the biomedical domain.
NLP Tasks
Named Entity Recognition (NER): Identify and classify mentions of cell lines or related entities appearing in biomedical contexts.
Relationship Extraction: Extract relationships between cell lines and other biomedical entities, such as genes, diseases, or drugs.
Text Classification: Classify sentences or articles based on their relevance to specific cell lines, particularly in cancer research or drug development.
Sentiment Analysis: Analyze the sentiment or tone of texts discussing cell lines, such as the evaluation of experimental results (positive or negative).
Information Retrieval: Develop systems to retrieve articles or specific mentions of cell lines based on user queries.
Entity Linking: Link cell line mentions in text to standardized identifiers in cell line ontologies or databases.
Question Answering (QA): Build systems that can answer specific questions about cell lines, such as their role in particular diseases or experiments.
Topic Modeling: Analyze the dataset to uncover major themes or trends in research involving cell lines.
Text Summarization: Automatically generate summaries of articles or sections discussing cell lines.
Who funded the creation of the dataset?
Any other comments?
The dataset aims to provide a foundational resource for advancing NLP in biomedicine.
Dataset Composition
- What experiments were initially run on this dataset? No experiments have been conducted yet. Updates will follow as they occur.
Data Collection Process
How was the data collected? The dataset was collected using the Europe PMC API. Articles marked as "open access" were retrieved, and those labeled as "retraction of publication" were excluded. Duplicate entries were filtered by ensuring unique PMCIDs.
Who was involved in the data collection process? The data collection process was carried out by researchers at EMBL-EBI, leveraging automated tools for querying and processing the Europe PMC repository.
Are there any known errors, sources of noise, or redundancies in the data? None have been identified yet.
Data Preprocessing
What preprocessing/cleaning was done?
- Only open-access articles were retrieved.
- Articles labeled as "retraction of publication" were excluded.
- Duplicate entries based on the PMCID column were removed.
- Paragraph text from each section of an article was Extracted with the relevant section referenced in the 'Section Column'
- Extra whitespace, inlne math/latex formatting and irrelevant sections such "Disclosure", "Publisher's note", etc, were filtered
- Name identifiers and personal data was also removed from the dataset
Dataset Distribution
How is the dataset distributed? The dataset is freely available for use and reproduction. Proper citation of the authors is required(Information to be updated).
When will the dataset be released/first distributed? To be updated.
Dataset Maintenance
Who is supporting/hosting/maintaining the dataset?
Europe PMC, and ChEMBL team responsible for the dataset's maintenance.
How does one contact the owner/curator/manager of the dataset?
Contact can be made via the community discussion forums on GitHub or Hugging Face.
Will the dataset be updated?
Yes, updates will occur as the project progresses.
How often and by whom?
Updates will be carried out periodically by the team.
How will updates/revisions be documented and communicated?
Updates will be documented via GitHub, using version tags.
Is there a repository to link to any/all papers/systems that use this dataset?
Yes, a GitHub repository will track publications and systems using this dataset.
If others want to extend/augment/build on this dataset, is there a mechanism for them to do so?
Yes, contributions are encouraged via GitHub. Quality will be assessed through pull requests, and accepted contributions will be communicated to users via version tags and release notes.
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