目的:探讨CRYAB与前列腺癌(PCa)预后的关系及可能机制。
方法:使用R软件进行生物信息学分析,包括差异基因表达和临床相关分析,接收机工作特性(ROC)曲线和Kaplan-Meier(KM)曲线生成。使用RT-qPCR检测基因表达,使用蛋白质印迹验证蛋白质表达。扩散,凋亡,用CCK8、TUNEL、Transwell迁移,和入侵检测。
结果:根据TCGA和GEO数据库,与正常组织相比,PCa组织中CRYABmRNA表达下调(P<0.05),与RWPE1细胞相比,PCa细胞中CRYABmRNA和蛋白表达下调(P<0.05)。细胞功能实验表明,上调CRYAB能够抑制细胞增殖,入侵,和前列腺癌细胞的迁移,促进细胞凋亡(P<0.05),并且在CRYAB上调的细胞系中上调CDH1表达同时下调CDH2表达。此外,CRYABmRNA表达与Gleason评分相关(P<0.01)。ROC曲线下面积为0.914,KM曲线显示CRYAB对无进展生存期(P=0.008)和疾病特异性生存期(P=0.032)具有预后价值。
结论:CRYAB在PCa组织中下调,与PCa细胞的抗肿瘤功能有关。它可能通过调节上皮-间质转化分子影响前列腺癌细胞的转移能力。CRYABmRNA在PCa中具有重要的诊断和预后价值。
OBJECTIVE: To explore the relationship between CRYAB and the prognosis of prostate cancer (PCa) as well as the potential mechanism.
METHODS: Bioinformatics analysis was performed using R software, including differential gene expression and clinical correlation analysis, receiver operating characteristic (ROC) curve and Kaplan-Meier (KM) curve generation. Gene expression was detected using RT-qPCR, and protein expression was validated using Western Blot. The proliferation, apoptosis, and metastatic ability of PCa cells were detected using CCK8, TUNEL, Transwell migration, and invasion assays.
RESULTS: According to the TCGA and GEO databases, CRYAB mRNA expression was down-regulated in PCa tissue compared with normal tissue (P< 0.05), and CRYAB mRNA and protein were down-regulated in PCa cells compared with RWPE1 cells (P< 0.05). Cell function experiments showed that up-regulated CRYAB could inhibit the proliferation, invasion, and migration of prostate cancer cells, promote apoptosis (P< 0.05), and up-regulate CDH1 expression while down-regulating CDH2 expression in the CRYAB-upregulated cell line. In addition, CRYAB mRNA expression was correlated with Gleason score (P< 0.01). The area under the ROC curve was 0.914, the KM curve showed that CRYAB had prognostic value for progression-free survival (P = 0.008) and disease-specific survival (P = 0.032).
CONCLUSIONS: CRYAB is down-regulated in PCa tissue and is associated with the anti- tumor function of PCa cells. It may affect the metastatic ability of prostate cancer cells by regulating epithelial-mesenchymal transition molecules. CRYAB mRNA has important diagnostic and prognostic value in PCa.