EphA2调控GPRC5A分子促进胰腺癌细胞增殖和迁移
首发时间:2024-04-22
摘要:目的:本研究旨在确定EphA2的下游靶点并探讨其调控机制,以更好地了解胰腺癌的侵袭性。方法:采用流式细胞术对EphA2高、低表达的PANC-1细胞进行筛选。用克隆形成实验检测EphA2对分选后细胞的增殖能力的影响。通过GEO数据库预测与EphA2相互作用的下游调控因子。采用实时定量聚合酶链反应(qRT-PCR)和蛋白免疫印迹法(WB)检测EphA2和GPRC5A的表达。采用Kaplan-Meier生存曲线分析TCGA数据库中胰腺癌组织中GPRC5A表达的临床意义。利用CRISPR-Cas9基因编辑技术敲除EphA2后探究胰腺癌细胞中GPRC5A表达的变化。Transwell实验检测EphA2低表达与过表达细胞间的迁移能力。结果:EphA2在胰腺癌细胞系中高表达,与胰腺癌细胞增殖能力呈正相关。随后筛选并验证了GPRC5A为EphA2的下游正向调控的效应蛋白。GPRC5A在临床样本中高表达,与预后不良、总生存率低相关。CRISPR-Cas9介导的EphA2敲除可以显著降低GPRC5A的表达水平,表明EphA2与GPRC5A呈正相关。此外,EphA2的过表达促进了胰腺癌细胞的迁移。结论:EphA2通过调控GPRC5A在促进胰腺癌细胞增殖中发挥重要作用,这可能为胰腺癌的治疗应用提供新的见解。
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EphA2 regulates the GPRC5A molecule enhancing cell proliferation and migration in pancreatic cancer
Abstract:Objective: The study aims to identify the downstream targets of EphA2 and explore the regulatory mechanisms to understand the invasiveness of pancreatic cancer better. Methods: PANC-1 cells with high or low EphA2 expression were sorted by flow cytometry. Colony formation assay was used to detect the proliferation function of EphA2 on the sorted cells. The downstream regulators that interacted with EphA2 were predicted by the GEO database. Real-time quantitative polymerase chain reaction (qRT-PCR) and Western blotting (WB) were used to detect the expression of EphA2 and GPRC5A. Kaplan-Meier analysis was used to analyze the clinical significance of GPRC5A expression in pancreatic cancer tissues in the TCGA database. CRISPR-Cas9 mediated knockout of EphA2 detected the change of GPRC5A expression in pancreatic cancer cells. Transwell assay detected the migration ability between EphA2 low and its overexpression cells. Results: EphA2 was highly expressed in pancreatic cancer cell lines and positively correlated with the proliferation of pancreatic cancer cells. GPRC5A was screened and validated as a downstream and positive effector protein of EphA2. GPRC5A showed high expression in clinical samples and was associated with poor prognosis and low overall survival. CRISPR-mediated knockout of EphA2 significantly reduced the expression level of GPRC5A, which indicated that EphA2 positively correlated to GPRC5A. In addition, overexpression of EphA2 promoted the migration of pancreatic cancer cells. Conclusion: EphA2 plays an important role in promoting the proliferation of pancreatic cancer cells by regulating GPRC5A. This may provide new insights into pancreatic cancer therapeutic applications.
Keywords: pancreatic cancer EphA2 GPRC5A proliferation migration
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EphA2调控GPRC5A分子促进胰腺癌细胞增殖和迁移
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