Department of Urology, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu 610072, China.
Department of Urology, West China Hospital of Sichuan University, Chengdu 610041, China.
Asian Pac J Trop Med. 2014 Jan;7(1):44-7. doi: 10.1016/S1995-7645(13)60190-X.
To study the expression of TRPC6 among prostate cancer cells, establish high expression cell lines of TRPC6, and to provide potential cell mode for prostate cancer oncogenesis and development.
Occurrence and development of prostate cancer cells, PC3, PC-3 m DU145, 22 rv1, LNCaP and normal prostate epithelial cells in the PrEC TRPC6 expression level were detected by QPCR method. Calcium phosphate transfection method was used to package retrovirus pLEGFP-N1-TRPC6 and pLEGFP-N1-vector and infect the prostate cancer cells, a stable high expression of TRPC6 prostate cancer cells. Sable cell lines of TRPC6, matrix metalloproteinase (MMP) 2, MMP9 expression was detected by QPCR and Western blot. Change of cell invasion ability was detected by Transwell.
The expression level of prostate cancer cells TRPC6 were higher than control group PrEC cells. Among TPRC6 the expression of cell line PC 3 transfer potential wre the lowest, and high transfer cell line PC-3M express was the highest. Real-time fluorescent quantitative PCR and western blot results showed that after filter, the seventh generation of cell TRPC6 protein and mRNA expression levels were higher than the control group obviously. Transwell experimental results showed that the overexpression of TRPC6 could promote the invasion ability of PC3 prostate cancer cells.
TRPC6 expressed in prostate cancer cells is in disorder, and its action may be associated with the invasion and metastasis of prostate cancer cells; successful establishment of stable high expression of TRPC6 prostate cancer cells primarily confirm the invasion-trigger ability of TRPC6 on prostate cancer, and lay down the Foundation for exploring the TRPC6's role in the occurrence and development of prostate cancer mechanism.
研究 TRPC6 在前列腺癌细胞中的表达，建立 TRPC6 高表达细胞系，为前列腺癌的发生发展提供潜在的细胞模型。
采用 QPCR 方法检测前列腺癌细胞 PC3、PC-3m、DU145、22rv1、LNCaP 及正常前列腺上皮细胞 PrEC 中 TRPC6 的表达水平。采用磷酸钙转染法包装逆转录病毒 pLEGFP-N1-TRPC6 和 pLEGFP-N1-载体并感染前列腺癌细胞，获得稳定高表达 TRPC6 的前列腺癌细胞系。采用 QPCR 和 Western blot 检测 TRPC6、基质金属蛋白酶（MMP）2、MMP9 的表达。采用 Transwell 检测细胞侵袭能力的变化。
前列腺癌细胞 TRPC6 的表达水平高于对照组 PrEC 细胞。在 TRPC6 中，PC 3 细胞系的转移潜能最低，而高转移细胞系 PC-3M 的表达最高。实时荧光定量 PCR 和 Western blot 结果显示，转染后第 7 代细胞 TRPC6 蛋白和 mRNA 表达水平明显高于对照组。Transwell 实验结果表明，TRPC6 的过表达可促进 PC3 前列腺癌细胞的侵袭能力。
前列腺癌细胞中 TRPC6 表达紊乱，其作用可能与前列腺癌细胞的侵袭和转移有关；成功建立稳定高表达 TRPC6 的前列腺癌细胞系，初步证实了 TRPC6 对前列腺癌的侵袭触发能力，为探讨 TRPC6 在前列腺癌发生发展中的作用机制奠定了基础。
