HRAS基因表达对宫颈癌细胞自噬及凋亡作用及机制  

ROLE AND MECHANISM OF HRAS GENE EXPRESSION IN THE AUTOPHAGY AND APOPTOSIS OF CERVICAL CANCER CELLS

作　　者：阎臻[1] 付晓瑞[2] 李新敏[3] 崔珺[1] YAN Zhen;FU Xiaorui;LI Xinmin;CUI Jun(Gynaecology And Obstetrics Department,Women&Infants Hospital of Zhengzhou,Zhengzhou 450052,China)

机构地区：[1]郑州市妇幼保健院妇产科,河南郑州450052 [2]郑州大学第一附属医院肿瘤科 [3]郑州市妇幼保健院病理科

出　　处：《青岛大学学报（医学版）》2021年第2期240-245,共6页Journal of Qingdao University(Medical Sciences)

基　　金：河南省医学科技攻关计划项目(201503211)。

摘　　要：目的探讨Harvery鼠肉瘤病毒癌基因(HRAS)表达对宫颈癌细胞自噬及凋亡的作用及其机制。方法构建宫颈癌小鼠模型,苏木精-伊红(HE)染色验证小鼠模型构建是否成功。将宫颈癌Hela细胞分为4组,空白对照组(A组:不转染任何序列)、阴性对照组(B组:转染空载体质粒)、HRAS vector组(C组:转染HRAS过表达质粒)、si-HRAS组(D组:转染HRAS沉默质粒siRNA)。实时定量聚合酶链反应(qRT-PCR)和Western blot分别检测宫颈癌组织和细胞中HRAS、磷酸酶和张力蛋白同源物基因(PTEN)、自噬相关基因7(ATG7)、微管相关蛋白1轻链3(LC3)、Beclin1、BCL2关联X蛋白(bax)、TNF受体超家族成员6(Fas)和E-cadherin的mRNA和蛋白表达水平,流式细胞术检测转染各组细胞凋亡情况。结果HE染色结果显示宫颈癌小鼠造模成功。宫颈癌小鼠模型中HRAS呈高表达,PTEN表达下调(t=9.450~14.260,P<0.05)。细胞转染后HRAS vector组HRAS、bcl-2 mRNA和蛋白表达水平明显上升,PTEN、ATG7、LC3、Beclin1、bax、Fas和E-cadherin均呈低表达,细胞凋亡率明显下降(F=1.011~220.400,P<0.05);而si-HRAS组趋势相反(F=3.160~622.800,P<0.05)。结论HRAS基因表达下调可能通过激活PTEN信号通路促进小鼠宫颈癌细胞自噬和凋亡,抑制上皮间质转化,从而抑制宫颈癌的发生。Objective To investigate the role and mechanism of V-Ha-Ras Harvey rat sarcoma viral oncogene homolog(HRAS)gene expression in the autophagy and apoptosis of cervical cancer cells.Methods A mouse model of cervical cancer was established,and HE staining was used to verify whether this model was successfully established.Cervical cancer Hela cells were divided into blank control group(no transfection of any sequences),negative control group(transfection with empty vector plasmid),HRAS vector group(transfection with HRAS overexpression plasmid),and si-HRAS group(transfection with HRAS-silencing plasmid siRNA).Quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression levels of HRAS,phosphatase and tensin homolog(PTEN),autophagy-related gene 7(ATG7),microtubule-associated protein 1 light chain 3(LC3),Beclin1,Bcl2-associated X protein(bax),tumor necrosis factor receptor superfamily member 6(Fas),and E-cadherin in cervical cancer tissue and cells,and flow cytometry was used to observe apoptosis in each group.Results HE staining showed that the mouse model of cervical cancer was successfully established.HRAS was highly expressed and PTEN expression was downregulated in the mouse model of cervical cancer(t=9.450-14.260,P<0.05).After transfection,the HRAS vector group had significant increases in the mRNA and protein expression of HRAS and Bcl-2,low expression of PTEN,ATG7,LC3,Beclin1,bax,Fas,and E-cadherin,and a significant reduction in cell apoptotic rate(F=1.011-220.400,P<0.05),while the si-HRAS group showed opposite trends(F=3.160-622.800,P<0.05).Conclusion By activating the PTEN signaling pathway,downregulation of the HRAS gene may promote the autophagy and apoptosis of cervical cancer cells in mice,inhibit epithelial-mesenchymal transformation,and thus inhibit the development of cervical cancer.

关 键 词：基因 ras 宫颈肿瘤 自噬性细胞死亡 细胞凋亡 PTEN信号通路 

分 类 号：R394.2[医药卫生—医学遗传学] R737.33[医药卫生—基础医学]