miR-21调控PI3K/Akt信号通路诱导宫颈癌细胞凋亡的机制研究

Mechanism of miR-21 regulating PI3K/Akt signaling pathway to induce apoptosis of cervical cancer cells

目的:观察miR-21对PI3K/Akt信号通路活化和宫颈癌细胞凋亡的影响,初步探讨相关作用机制。方法:利用miRNA表达数据库和TCGA生存分析数据库分析miRNA与宫颈癌的关系。选取宫颈癌患者63例和妇科体检正常人60例作为研究对象,分别用实时荧光定量聚合酶链反应(qPCR)法和蛋白质印迹(Western blot)法检测患者宫颈癌组织、癌旁组织以及正常宫颈组织miR-21、PI3K、Akt蛋白相对表达量。将人宫颈癌Hela细胞分为NC组(转染阴性对照序列)、mimics组(转染miR-21 mimics)和inhibitor组(转染miR-21 inhibitor),qPCR检测转染细胞miR-21、PI3K mRNA和Akt mRNA相对表达量,Western blot检测转染细胞PI3K和Akt蛋白表达水平,流式细胞术检测细胞凋亡情况。结果:生物信息学分析方法发现,miR-21低表达的患者有良好的预后(P<0.05)。宫颈癌组织miR-21相对表达量明显高于癌旁组织和正常组织,PI3K m RNA、Akt mRNA及蛋白相对表达量明显高于癌旁组织和正常组织,且癌旁组织高于正常组织(均P<0.05)。转染后,mimics组细胞凋亡率低于NC组,inhibitor组细胞凋亡率高于NC组(均P<0.05);mimics组p-PI3K、PI3K、p-Akt、Akt蛋白及其mRNA表达水平均高于NC组和inhibitor组(P<0.05),inhibitor组低于NC组(P<0.05);mimics组p-PI3K/PI3K和p-Akt/Akt比值高于NC组和inhibitor组,inhibitor组低于NC组(P<0.05)。结论:宫颈癌发病与miR-21过表达相关,miR-21可能通过激活PI3K/Akt信号通路抑制宫颈癌细胞凋亡。

Objective:To observe the effects of miR-21 on the activation of PI3 K/Akt signaling pathway and apoptosis of cervical cancer cells,and to study the related mechanism.Methods:miRNA expression database and TCGA survival analysis database were used to analyze the relationship between miRNA and cervical cancer.Sixty-three cervical cancer patients and 60 normal people from gynecological examination were enrolled as the research subjects.Real-time fluorescent quantitative polymerase chain reaction(qPCR)and Western blot were used to detect the miR-21,PI3 K,and Akt mRNA and proteins level in cervical cancer tissues,paracancerous tissues,and normal cervical tissues.Human cervical cancer Hela cells were divided into NC group(transfected with negative control sequence),mimics group(transfected with miR-21 mimics),and inhibitor group(transfected with miR-21 inhibitor).qPCR was used to detect the relative expression of miR-21,PI3 K mRNA and Akt mRNA in transfected cells,Western blot was used to detect expression levels of PI3 K and Akt protein,and flow cytometry was used to detect cell apoptosis.Results:Bioinformatics analysis showed that patients with low expression of miR-21 had a good prognosis(P<0.05).The relative expression of miR-21 in cervical cancer tissues was significantly higher than that in adjacent tissues and normal tissues.The relative expression level of mRNA and protein of PI3 K and Akt in cervical cancer tissues were significantly higher than that in adjacent tissues and normal tissues,and they were higher in the adjacent tissues than that in normal tissues(all P<0.05).After transfection,the apoptosis rate of mimics group was lower than that of NC group,and the apoptosis rate of inhibitor group was higher than that of NC group(all P<0.05).The expression levels of mRNA and protein of p-PI3 K,PI3 K,p-Akt,and Akt were higher in mimics group than that in NC group and inhibitor group(P<0.05),and they were lower in inhibitor group than that in NC group(P<0.05).The ratios of p-PI3 K/PI3 K and p-Akt/Akt were higher in mimics group than that in NC group and inhibitor group,and they were lower in inhibitor group than that in NC group(P<0.05).Conclusion:Cervical cancer is associated with miR-21 overexpression,miR-21 may inhibit the apoptosis of cervical cancer cells by activating the PI3 K/Akt signaling pathway.