RT-PCR Array技术筛选膀胱癌癌症相关信号通路基因的表达变化

The expression changes of cancer-related pathways genes screened by RT-PCR Array in bladder cancer

背景与目的:膀胱癌是泌尿外科最常见肿瘤疾病,其发病机制至今尚未完全明了。本研究旨在观察正常膀胱组织与膀胱癌组织众多肿瘤相关信号通路关键基因的表达情况,为后续进一步深入研究膀胱癌复发与转移提供证据。方法:收集27例膀胱癌患者标本,液氮保存,冰冻切片法分离膀胱癌组织与正常膀胱组织,用德国QIAGEN公司Cancer Pathway Finder PCR Array板筛选与癌症相关信号通路中的84个基因在膀胱癌组织和正常膀胱组织中的表达情况。结果:与正常膀胱组织相比,膀胱癌组织中上调基因8个,下调基因19个。本研究选择影响上皮细胞间质化(epithelial-mesenchymal transition,EMT)的信号通路作为研究方向,其中GSC、KRT14和DSP上调,SNAI2和SNAI3下调。因此,选定GSC、KRT14、DSP、SNAI2和SNAI3作为兴趣基因,并对其进行多样本荧光定量q RT-PCR验证。结果表明:GSC在膀胱癌组织中上调,但与正常膀胱组织表达相比,差异无统计学意义(P>0.05)。KRT14和DSP在膀胱癌中表达高于正常组织(P<0.05),SNAI2和SNAI3在膀胱癌中表达低于正常膀胱组织(P<0.05),且SNAI3表达差异最为明显。结论:基因KRT14、DSP及SNAI3可能与膀胱癌发生、发展及转移有密切关系,可能成为基因治疗膀胱癌的重要靶点。

Background and purpose： Bladder cancer is the most common urological tumor, and its pathogenesis is still not fully understood. The study was aimed to observe the expressions of key genes in many tumor-associated signaling pathways in normal bladder tissue and bladder carcinoma, and to provide further evidence for the subsequent study of bladder cancer recurrence and metastasis. Methods： Twenty-seven eases of bladder cancer specimens were collected, and normal bladder tissues and bladder cancer tissues were distinguished by frozen section. Then, the expressions of 84 genes of cancer-related signaling pathways in bladder cancer tissues and normal bladder tissues were screened by Cancer Pathway Finder PCR Array produced by QIAGEN company. Results： Compared with the normal bladder tissues, the bladder carcinoma tissues had 8 up-regulated genes and 19 down-regulated genes. In this study, the impact of epithelial-mesenchymal transition （EMT） signaling pathway was selected as a research direction in which the GSC, KRT14, DSP were up-regulated, SNA12, SNA13 were down-regulated. Therefore GSC, KRT14, DSP, SNA12 and SNA13 were chosen as target genes, and verified by qRT-PCR in many examples. The result showed that the expressions of GSC gene in bladder cancer tissues were up-regulated, but with no statistical significance; KRT14, DSP expressions in bladder cancer were higher than those in normal bladder tissues （P〈0.05）; SNA12, SNA13 expressions in bladder cancer were lower than those in normal bladder tissues （P〈0.05）, and SNA13 showed the most obvious expression differences. Conclusion： KRT14, DSP and SNA13 may play an important role in bladder cancer＇s occurrence, development and metastasis.