SIRT1调控Nrf2信号通路对宫颈癌细胞增殖、凋亡及迁移侵袭的影响

Effects of SIRT1 on Nrf2 signaling pathway on proliferation, apoptosis, migration and invasion of cervical cancer cells

目的探讨SIRT1调控Nrf2信号通路对宫颈癌细胞增殖、凋亡及迁移侵袭的影响。方法选取宫颈癌细胞株Hela细胞,将Hela细胞株分为:空白对照组、SIRT1阴性对照组(negative control group,NC组)和SIRT1抑制组;采用Western Blot和qRT-PCR法检测各组细胞SIRT1的表达水平;采用MTT法检测各组细胞的增殖能力;采用流式细胞术检测各组细胞的凋亡能力;采用划痕实验和Transwell侵袭实验检测细胞迁移、侵袭能力;采用Western Blot和qRT-PCR法检测各组细胞Nrf2信号通路相关基因的表达水平。结果转染SIRT1干扰载体特异性抑制Hela细胞中SIRT1基因的表达,Western Blot和qRT-PCR结果均显示,SIRT1抑制组Hela细胞中SIRT1基因的蛋白和mRNA表达水平均低于空白对照组和SIRT1 NC组(均P<0.05)。MTT结果显示,48 h、96 h后SIRT1抑制组细胞的吸光度值均低于空白对照组和SIRT1 NC组(P<0.05)。细胞凋亡结果显示,与空白对照组和SIRT1 NC组相比,SIRT1抑制组细胞的凋亡能力显著增加(P<0.05);细胞周期结果发现,SIRT1抑制组G0/G1期细胞数明显多于空白对照组和SIRT1 NC组;而SIRT1抑制组S期细胞数明显减少(P<0.05)。细胞划痕和Transwell细胞侵袭实验结果显示,SIRT1抑制组划痕宽度较空白对照组和SIRT1 NC组更宽、穿过小室基质胶的细胞数目显著少于空白对照组和SIRT1 NC组(P<0.05)。与空白对照组和SIRT1 NC组相比,SIRT1抑制组Nrf2、NQO1、HO-1的蛋白及mRNA表达水平均显著降低(均P<0.05)。结论SIRT1可促进宫颈癌细胞增殖、迁移和侵袭,抑制其凋亡,其可能的作用机制是SIRT1促进Nrf2信号通路的活化。

Objective To investigate the effect of SIRT1 on Nrf2 signaling pathway on the proliferation,apoptosis,migration and invasion of cervical cancer cells.Methods Hela cell line was selected and divided into three groups:blank control group,SIRT 1 NC group and SIRT1 inhibition group.Western Blot and qRT-PCR were used to detect SIRT1 expression in each group.MTT method was used to detect the proliferation of cells in each group,flow cytometry was used to detect the apoptosis of cells in each group,scratch test and Transwell invasion test were used to detect the migration and invasion of cells.Western Blot and qRT-PCR were used to detect the expression level of Nrf2 signal pathway related genes.Results Western Blot and qRT-PCR results showed that SIRT1 protein and mRNA expression levels in Hela cells of SIRT1 inhibition group were lower than those of blank control group and SIRT1 NC group(all P<0.05).The results of MTT showed that the absorbance of SIRT 1 inhibition group were lower than those of blank control group and SIRT 1 NC group after 48 h and 96 h(P<0.05).Compared with the blank control group and SIRT 1 NC group,the apoptosis ability of SIRT 1 inhibition group was significantly increased(P<0.05);The cell cycle results showed that the number of G0/G1 phase cells in SIRT1 inhibition group was significantly higher than that in the blank control group and SIRT1 NC group;While the number of S phase cells in SIRT1 inhibition group was significantly reduced(P<0.05).The results of cell scratch and Transwell cell invasion showed that the scratch width of SIRT1 inhibition group was wider than that of the blank control group and SIRT1 NC group,and the number of cells passing through the chamber matrix glue in SIRT1 inhibition group were significantly less than those in the blank control group and SIRT1 NC group(P<0.05).Compared with the blank control group and SIRT1 NC group,the protein and mRNA expression levels of Nrf2,NQO1 and HO-1 in SIRT1 inhibition group were significantly reduced(all P<0.05).Conclusion SIRT1 can promote the proliferation,migration,invasion and inhibit the apoptosis of cervical cancer cells.The possible mechanism is that SIRT 1 can promote the activation of Nrf 2 signaling pathway.