摘要
目的研究VEGF-C对体外培养的宫颈癌Hela细胞增殖和凋亡的分子机制。方法应用重组人VEGF-C蛋白体外刺激宫颈癌Hela细胞,MTT法检测细胞增殖、流式细胞仪检测细胞周期和凋亡、Western Blotting检测增殖凋亡相关基因Bcl-2、cyclin D1蛋白水平的变化。结果 10、20、50 ng/μl VEGF-C处理后,细胞增殖指数分别为(1.00±0.03)、(1.25±0.05)、(1.55±0.08)、(2.13±0.08),呈剂量依赖性升高(P<0.05)。细胞周期S期比率呈剂量依赖性升高(P<0.05),分别为(30.91±0.09)%、(37.95±0.27)%、(45.05±0.40)%、(64.26±0.20)%;细胞凋亡率呈剂量依赖性降低(P<0.05),分别为(12.4±0.3)、(11.4±0.2)、(9.6±0.15)、(5.5±0.25)。Bcl-2、cyclin D1蛋白表达呈剂量依赖性升高。结论外源性VEGF-C通过细胞内信号的传递,诱导cyclin D1的表达,使肿瘤细胞S期加快,促进细胞周期的进程,来促进Hela细胞增殖;诱导Bcl-2的表达,抑制凋亡。
Objective To investigate the molecular mechanisms of Cell proliferation and apoptosis induced by VEGF - C in hu- man cervical cancer Hela cells. Methods Hela cells were stimulated by recombinant VEGF - C in vitro. Cell proliferation was detected by M3~I＂ assay. Cell cycle and cell apoptosis were detected by flow eytometry （FCM）. The expression of prolifera- tion and apoptosis related genes, Bcl - 2 and cyclin D1, were determined by western blotting. Results VEGF- C of 10 ng/ μl, 20 ng/μl and 50 ng/μl was found to increase the cell proliferation to 1.25± 0.05,1.55 ± 0.08 and 2.13 ± 0.08 when com- pared to the control 1.00 ± 0.03. And the cell number in S phase was increased to （37.95 ± 0.27） %, （45.05 ± 0.40） % and （64.26 ± 0.20） % from the control （30.91 ± 0.09） %. The cell apoptosis was inhibited in a dose - dependent manner, with the apoptosis index decreasing to 11.4 ± 0.2, 9.6 ± 0.15 and 5.5 ± 0.25 from the control 12.4 ± 0.3 （P 〈 0.05）. The expression levels of Bcl - 2 and cyclin D1 were increased in a dose dependent manner after being stimulated by VEGF - C （P〈0.05）. Conclusions Exogenous VEGF- C induces the expression of cyciin D1, which accelerates cell cycle phase S and promotes cell proliferation. It also inhibits cell apoptosis by inducing the expression of Bel- 2.
出处
《实用预防医学》
CAS
2014年第2期135-139,共5页
Practical Preventive Medicine
基金
国家自然科学基金面上项目(项目编号81201871)
