miR-125b靶向Fas/FasL信号传导途径影响胃癌细胞增殖  

MiR-125b inhibits growth of gastric cancer cells by targeting Fas/FasL signaling pathway

作　　者：李忆东 李婷婷[1] 赵珠峰[1] 吴国忠 Li Yidong;Li Tingting;Zhao Zhufeng;Wu Guozhong(Department of Basic Medicine,International School of Medical Technology,Shanghai Shanda University,Shanghai 201209,China)

机构地区：[1]上海杉达学院国际医学技术学院基础医学部,上海201209

出　　处：《中国医师杂志》2022年第6期854-858,共5页Journal of Chinese Physician

基　　金：国家自然科学基金(81172100);上海杉达学院护理学重点学科重点立项课题(2019zz35-yj)。

摘　　要：目的探究miR-125b在凋亡相关蛋白(Fas)/凋亡相关蛋白配体(FasL)信号介导胃癌细胞生长中的调控作用。方法体外培养胃癌SGC-7901细胞,将miR-125b inhibitor序列、NC序列、转染试剂分别转染至SGC-7901细胞中,分为miR-125b抑制组、NC组、Control组。用实时荧光定量PCR(qRT-PCR)法检测转染后细胞miR-125b表达,CCK-8法检测细胞增殖情况;平板细胞克隆形成实验检测菌落形成情况;流式细胞术检测细胞凋亡以及周期情况;蛋白免疫印迹法(Western blot)检测Fas、FasL的蛋白表达;荧光素酶活性测定检测miR-125b对Fas的靶向调控作用。结果miR-125b抑制组细胞miR-125b表达水平及细胞菌落形成数量显著低于Control组、NC组,细胞增殖抑制率、凋亡率显著高于Control组、NC组(均P<0.05)。miR-125b抑制组G1期DNA含量显著高于Control组、NC组,S期细胞DNA含量显著低于Control组、NC组(均P<0.05)。miR-125b抑制组Fas、FasL蛋白表达显著高于Control组、NC组(均P<0.05)。Fas mRNA的3′-UTR中存在miR-125b的靶位点,与NC+Fas 3′UTR-Wt组相比,miR-125b抑制组+Fas 3′-UTR-Wt组荧光素酶活性显著降低(P<0.05)。结论抑制miR-125b表达后可激活Fas/FasL信号进而抑制SGC-7901细胞增殖,引起细胞周期G1期阻滞,促进其凋亡。Objective To investigate the regulation effect of miR-125b in the gastric cancer cell growth mediated by apoptosis related protein(Fas)/apoptosis related protein ligand(FasL)signal.Methods Gastric cancer SGC-7901 cells were cultured in vitro.MiR-125b inhibitor sequence,NC sequence and transfection reagent were transfected into SGC-7901 cells and divided into three groups:miR-125b inhibited group,NC group and control group.The expression of miR-125b in transfected cells was detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR),and cell proliferation was detected by cell counting kit-8(CCK-8)method.The colony formation was detected by plate cell clone formation assay.Cell apoptosis and cycle were detected by flow cytometry.The protein expression of Fas and FasL was detected by Western blot.The targeted regulation of Fas by miR-125b was detected by luciferase activity assay.Results The expression level of miR-125 and the number of cell colony in miR-125b inhibited group was significantly lower than those in control group and NC group,and the inhibition rate of cell proliferation and apoptosis rate were significantly higher than that in control group and NC group(all P<0.05).The DNA content in G1 phase in miR-125b inhibited group was significantly higher than that in control group and NC group,and the DNA content in S phase in miR-125b inhibited group was significantly lower than that in control group and NC group(all P<0.05).The expression of Fas and FasL protein in miR-125b inhibited group was significantly higher than that in control group and NC group(all P<0.05).The target site of miR-125b was found in 3'-UTR of Fas mRNA,and compared with the NC+Fas 3'UTR-Wt group,the activity of luciferase in the miR-125b inhibited group+Fas 3'-UTR-Wt group decreased significantly(P<0.05).Conclusions Inhibition of miR-125b expression can activate Fas/FasL signal and inhibit SGC-7901 cell proliferation,induce G1 phase arrest of cell cycle and promote apoptosis.

关 键 词：胃肿瘤 细胞系 肿瘤 miR-125b Fas相关死亡结构域蛋白质 Fas配体蛋白质 

分 类 号：R735.2[医药卫生—肿瘤]