GPRC6A过表达前列腺癌细胞株构建及EMT相关研究

发布时间：2018-01-31 13:25

本文关键词： GPRCA 前列腺癌上皮间质转化　出处：《中国生物工程杂志》2017年03期 　论文类型：期刊论文

【摘要】：目的:构建GPRC6A基因过表达的前列腺癌LncapC4-2细胞株,检测细胞迁移和侵袭能力改变及EMT相关基因表达,为研究GPRC6A介导的EMT在前列腺癌发生发展过程中的作用奠定基础。方法:构建过表达GPRC6A的慢病毒载体pCDH-GPRC6A,人胚肾293T细胞包被病毒并感染Lncap C4-2细胞株,Puromycin筛选获得稳定过表达GPRC6A的细胞株。RT-PCR和Western blot验证细胞GPRC6A的过表达情况。划痕和transwell实验检测细胞迁移和侵袭能力,qPCR检测EMT相关基因表达。结果:成功构建GPRC6A表达的慢病毒载体,RT-PCR和Western blot检测LncapC4-2 GPRC6A过表达细胞株中GPRC6A的mRNA和蛋白质表达增高,与对照相比较,在过表达GPRC6A的LncapC4-2细胞中,细胞迁移和侵袭能力增强,并且EMT相关基因E-cadherin表达降低而SNAIL表达增高。结论:成功构建过表达的GPRC6A的前列腺癌细胞株,肿瘤细胞中的GPRC6A可能通过增强细胞迁移和侵袭能力并促进细胞EMT而参与前列腺癌发生发展过程。
[Abstract]:Objective: to construct a prostate cancer LncapC4-2 cell line with overexpression of GPRC6A gene and to detect the change of migration and invasion ability and the expression of EMT related genes. To study the role of EMT mediated by GPRC6A in the development of prostate cancer. Methods: a lentivirus vector pCDH-GPRC6A was constructed to express GPRC6A. Human embryonic kidney 293T cells were coated with virus and infected with Lncap C4-2 cell line. Puromycin screening of cells with stable overexpression of GPRC6A. RT-PCR and Western. Blot was used to verify the overexpression of GPRC6A. Scratch and transwell assay were used to detect cell migration and invasion. QPCR was used to detect the expression of EMT related genes. Results: the lentivirus vector expressing GPRC6A was successfully constructed. RT-PCR and Western blot were used to detect the increase of mRNA and protein expression of GPRC6A in LncapC4-2 GPRC6A overexpression cell lines. Compared with the control group, the migration and invasion of LncapC4-2 cells with overexpression of GPRC6A were enhanced. The expression of E-cadherin was decreased and the expression of SNAIL was increased. Conclusion: the prostate cancer cell line with overexpression of GPRC6A was successfully constructed. GPRC6A in tumor cells may be involved in the development of prostate cancer by enhancing cell migration and invasion and promoting cellular EMT.
【作者单位】：山西医科大学基础医学院;北京协和医学院研究生院;
【基金】：国家青年科学基金(81302220) 山西省青年科学基金(2014021037-1) 山西医科大学博士启动基金(B03201204)资助项目
【分类号】：R737.25
【正文快照】： 近年来,随着我国老龄化人口的增长,前列腺癌正成为严重影响我国男性健康的泌尿系恶性肿瘤[1-2]。前列腺癌好发骨转移,加重病情的发展[3]。有必要寻找前列腺癌骨转移的标志物,为前列腺癌骨转移有效防治提供新思路和药物作用的新靶点。GPRC6A是2004年发现的G蛋白耦联受体C家族成

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