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目的探讨人类精液通过细胞外信号调节激酶(ERK)通路对宫颈癌He La细胞的增殖、凋亡及迁移的影响。方法用不同浓度的人类精液刺激He La细胞,MTT法分析精液对He La细胞的促增殖作用,Annexin V-FITC/P法检测各组细胞凋亡的影响,PI法检测各组细胞周期的变化,Transwell小室体外细胞侵袭实验检测对He La细胞侵袭能力的影响,蛋白质印迹实验检测ERK信号通路相关分子的表达情况,U0126抑制ERK信号通路检测精液诱导的ERK1/2的磷酸化及c-myc的表达情况,同时检测精液对细胞增殖的影响。结果精液能促进He La细胞增殖,ERK1/2的磷酸化水平及c-myc的表达随精液浓度的升高而增加,U0126能抑制精液诱导的ERK1/2的磷酸化、c-myc的上调及细胞的增殖,而与对照组相比精液对宫颈癌细胞的凋亡率没有显著影响(P>0.05)。随着精液浓度的增加,宫颈癌细胞周期中S期和G2/M期细胞百率逐渐增高,尤其是1:10浓度组结果更为明显(P<0.05)。与对照组比较,随着精液浓度的增加穿膜细胞数明显增加,当精液的浓度为1∶50时差异开始有统计学意义(P<0.05)。当精液的浓度为1∶100时即能在细胞内检测到ERK1/2的磷酸化有升高的趋势,而随着精液浓度的增加ERK1/2的磷酸化显著上调,其下游靶基因c-myc的表达也显著上调(P<0.05)。结论精液能够通过活化ERK通路促进He La细胞增殖,并导致宫颈癌细胞周期中S期及G2/M期延长,促进宫颈癌细胞的侵袭能力增强。
Objective To investigate the effects of human seminal fluid on the proliferation, apoptosis and migration of Hela cells by extracellular signal-regulated kinase (ERK) pathway. Methods He La cells were stimulated with different concentrations of human semen. The proliferation of Hela cells was analyzed by MTT assay. The apoptosis of Hela cells was detected by Annexin V-FITC / P assay. The cell cycle was detected by PI assay. , Transwell cell invasion assay in vitro, the effect of ERK1 / 2 phosphorylation and c-myc on the activation of HeLa cell were detected by Western blotting. The expression of ERK signaling pathway-related molecules was detected by Western blotting. Expression, while testing the impact of semen on cell proliferation. Results Semen could promote the proliferation of HeLa cells. The phosphorylation of ERK1 / 2 and the expression of c-myc increased with the increase of seminal fluid concentration. U0126 inhibited the phosphorylation of ERK1 / 2 and the up-regulation of c-myc, Compared with the control group, seminal fluid had no significant effect on the apoptosis rate of cervical cancer cells (P> 0.05). With the increase of seminal fluid concentration, the percentage of cells in S phase and G2 / M phase of cervical cancer cell cycle gradually increased, especially in the 1:10 concentration group (P <0.05). Compared with the control group, the number of transmembrane cells increased significantly with the increase of seminal fluid concentration. When the semen concentration was 1:50, the difference began to have statistical significance (P <0.05). When the concentration of semen was 1: 100, the phosphorylation of ERK1 / 2 was detected in the cells with a rising trend. With the increase of semen concentration, the phosphorylation of ERK1 / 2 was significantly up-regulated. The downstream target gene c- The expression of myc was also significantly up-regulated (P <0.05). Conclusion Semen can promote the proliferation of HeLa cells via activation of ERK pathway and lead to the extension of S phase and G2 / M phase in cervical cancer cell cycle and promote the invasion of cervical cancer cells.