线粒体分裂融合基因在氧化应激诱导宫颈癌Hela细胞损伤中的作用

Effect of mitochondrial fusion and fission genes on cervical carcinoma Hela cells injury induced by oxydative stress

目的:利用维生素K3(VitaminK3,Vk3)复制子宫颈癌Hela细胞损伤模型,观察线粒体融合分裂基因的变化,探讨线粒体融合分裂基因在Vk3诱导Hela细胞凋亡过程中的作用。方法:MTT法检测各组Hela细胞存活率,Hoechst33258染色观察细胞凋亡,RT-PCR方法检测各组Hela细胞中线粒体融合基因Mfn1、Mfn2和Opa1及线粒体分裂基因Drp1、Fist1和MTP18mRNA表达。结果:Vk3能够降低Hela细胞存活率,并且Hoechst33258染色结果显示细胞呈现凋亡形态变化,凋亡率增加,Mfn1、Opa1、和MTP18mRNA表达量明显下降(P<0.05);而与Vk3单独作用组比较,Vk3+NAC联合作用组Hela细胞存活率增加(P<0.05),凋亡率降低,Mfn1、Opa1、和MTP18mRNA表达量明显增加(P<0.05)。结论:线粒体融合和分裂基因表达的降低都有可能参与了氧化应激诱导的细胞损伤作用。

Objective ： To establish injury model of cervical carcinoma Hela cells by vitamin K3, observe the changes of mitochondrial fusion and fission genes, explore the effect of mitochondrial fusion and fission genes on apoptosis of Hela cells induced by vitamin K3. Methods ： The survival rate of Hela cells was detected by MTT method, the apoptotis of Hela cells was observed by Hoechst33258 staining, the expression levels of Mfnl, Mfn2, Opal, Drpl, Fist1 and MTP18 mRNA were detected by RT - PCR. Results： Vitamin K3 reduced the survival rate of Hela cells, the results of Hoechst33258 staining showed that morphological changes of Hela cells appeared, apoptosis rate increased, the expression levels of Mfnl, Opal and MTP18 mRNA decreased （P 〈 0. 05 ） ; compared with vitamin K3 group, the survival rate of Hela ceils in vitamin K3 ＋ NAC group increased （ P 〈 0. 05 ）, the apoptosis rate decreased, the expression levels of Mfnl, Opal and MTP18 mRNA increased （ P 〈 0. 05 ） . Conclusion ： Downregulation of mitochondrial fusion and fission genes may be involved in the process of cell injury induced by oxydative stress.