微小RNA-107调控内质网应激对喉癌Hep-2细胞癌增殖及凋亡的影响  

Effect of miR-107 on the proliferation and apoptosis of laryngeal carcinoma Hep-2 cells by regulating endoplasmic reticulum stress

作　　者：杨雪[1] 冯志星[1] 马海滨[1] 任雪燕 单珊 韩海平[1] YANG Xue;FENG Zhixing;MA Haibin;REN Xueyan;SHAN Shan;HAN Haiping(Department of Otolaryngological,Handan Central Hospital of Hebei,Handan,Hebei 056001,China)

机构地区：[1]邯郸市中心医院耳鼻喉科,河北邯郸056001

出　　处：《安徽医药》2022年第5期904-907,共4页Anhui Medical and Pharmaceutical Journal

基　　金：河北省医学科学研究项目(20200485)。

摘　　要：目的探讨微小RNA(miR-107)调控内质网应激对喉癌Hep-2细胞增殖和凋亡的影响。方法将miR-107类似物和阴性对照片段转染至对应组细胞中,以未转染组作为空白组。采用荧光定量PCR(RT-PCR)检测各组Hep-2细胞中miR-107相对表达量,用蛋白质印迹法(Westernblotting)检测各组Hep-2细胞中的内质网应激有关蛋白即蛋白激酶R样内质网激酶(PERK)及活化转录因子4(ATF4)蛋白的表达,采用四甲基偶氮唑盐(MTT)比色法测定Hep-2细胞的增殖能力,用流式细胞术检测细胞凋亡情况。结果三组转染细胞miR-107相对表达量差异有统计学意义(χ^(2)=11.50,P<0.05)。其中mimics组miR-107的相对表达量为(15.32±1.94),与mimics组相比较,NG组、空白组miR-107相对表达量均降低(P<0.05)。三组转染细胞的PERK和ATF4蛋白表达水平均差异有统计学意义(F=70.35、13.77,P<0.05)。其中mimics组PERK和ATF4蛋白的相对表达量为(0.53±0.07)和(0.75±0.06),与mimics组相比较,NG组和空白组的PERK和ATF4蛋白相对表达量均降低(P<0.05)。MTT分析结果显示,三组细胞的增殖能力差异有统计学意义(F=21.44、28.38、59.14,P<0.05)。其中mimics组Hep-2细胞增殖能力在12、24、36 h时分别为(0.15±0.01)、(0.33±0.03)和(0.47±0.04),与mimics组比较,在12、24、36 h后NG组、空白组Hep-2细胞增殖能力均有显著升高(P<0.05)。三组转染细胞的细胞凋亡率比较差异有统计学意义(F=102.32,P<0.05)。其中mimics组Hep-2细胞凋亡率为(10.18±0.93),与mimics组相比较,NG组和空白组的细胞凋亡率均降低(P<0.05)。结论上调miR-107的表达能够抑制Hep-2细胞的增殖,促进Hep-2细胞凋亡,内质网应激可能在其中有重要作用。Objective To investigate the effect of miR-107 on the proliferation and apoptosis of laryngeal carcinoma Hep-2 cells by regulating endoplasmic reticulum stress.Methods The miR-107 analog and negative control fragment were transfected into the corresponding group of cells,and the untransfected group was used as the blank group.Fluorescence quantitative PCR(RT-PCR)was used to detect the relative expression of miR-107 in Hep-2 cells in each group,and Western blot(WB)was used to detect the expression of endoplasmic reticulum stress proteins including protein kinase R-like ER kinase(PERK)and Activating Transcription Factor 4(ATF4)protein in Hep-2 cells in each group.Tetramethylazolyzole(MTT)Chromatography was used to determine the proliferation ability of Hep-2 cells,and flow cytometry was used to detect cell apoptosis.Results The relative expression of miR-107 in the three groups of transfected cells was significantly different(χ^(2)=11.50,P<0.05).The relative expression of miR-107 in the mimics group was(15.32±1.94).Compared with the mimics group,the relative expression of miR-107 in the NG group and the blank group was decreased(P<0.05).The differences in the expression levels of PERK and ATF4 proteins of the three groups of transfected cells were statistically significant(F=70.35,13.77,P<0.05).The relative expression levels of PERK and ATF4 protein in the mimics group were(0.53±0.07)and(0.75±0.06).Compared with the mimics group,the relative expression levels of PERK and ATF4 proteins in the NG group and the blank group were both decreased(P<0.05).The results of MTT analysis showed that there was a statistically significant difference in the proliferation ability of the three groups of cells(F=21.44,28.38,59.14,P<0.05).The proliferation ability of Hep-2 cells in the mimics group was(0.15±0.01),(0.33±0.03)and(0.47±0.04)at 12,24,and 36 h,respectively.Compared with the mimics group,the proliferation ability of Hep-2 cells in the NG group and the blank group was significantly increased after 12,24,and 36 h(P<0.0

关 键 词：微小RNA-107 内质网应激 喉癌HEP-2细胞 增殖 凋亡 

分 类 号：R739.65[医药卫生—肿瘤]