下调TMEM97对卵巢癌细胞SKOV3增殖、凋亡的影响及其相关机制探讨  

Effects and mechanism of down-regulated TMEM97 on proliferation and apoptosis of ovarian cancer SKOV3 cells

作　　者：刘晋芳 徐小燕[2] 周儒奎 徐慧超 高艳 苗宇船 LIU Jin-fang;XU Xiao-yan;ZHOU Ru-kui;XU Hui-chao;GAO Yan;MIAO Yu-chuan(Department of Basic Medical Sciences,Basic Medical College,Shanxi University of Traditional Chinese Medicine,Taiyuan 030024,China;Department of Pathophysiology,Basic Medical College,China Medical University;The Graduate School,Shanxi University of Traditional Chinese Medicine)

机构地区：[1]山西中医药大学基础医学院基础医学系,030024 [2]中国医科大学基础医学院病理生理学教研室 [3]山西中医药大学研究生院

出　　处：《天津医药》2020年第8期695-699,共5页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(81470190);山西中医药大学科技创新能力培育计划项目(2019PY-026)。

摘　　要：目的观察下调跨膜蛋白97(TMEM97)对卵巢癌细胞SKOV3增殖和凋亡的影响,并探讨其相关机制。方法将卵巢癌细胞SKOV3分为转染组(si-TMEM97组)、阴性对照组(siNC组)和空白对照组(Control组)。采用实时荧光定量PCR(qPCR)及Western blot法分别在mRNA和蛋白表达水平评估转染siRNA-TMEM97后TMEM97的沉默效果;分别在转染后采用CCK-8法、PI染色法、Annexin V-FITC/PI染色法和流式细胞术检测下调TMEM97对细胞的增殖、细胞周期和细胞凋亡的影响,并检测B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白质(Bax)及P38丝裂原活化蛋白激酶(P38 MAPK)通路相关蛋白(P38/MAPK、p-P38/MAPK)的表达变化。结果si-TMEM97组细胞的增殖能力相比Control组和siNC组降低,而早期凋亡率增高(均P<0.01)。si-TMEM97组细胞Bcl-2的蛋白相对表达量较Control组和siNC组降低,而Bax、p-P38/MAPK的蛋白相对表达量较Control组和siNC组增高(均P<0.05)。结论下调TMEM97的表达使卵巢癌细胞SKOV3增殖减少,凋亡增加,这可能与调节凋亡相关蛋白Bcl-2/Bax表达及P38/MAPK信号通路的激活有关。Objective To explore the effects of transmembraneprotein 97(TMEM97)on the proliferation and apoptosis of human ovarian cancer SKOV3 cells and its mechanism.Methods The SKOV3 cells were divided into transfection(si-TMEM97)group,negative control(siNC)group and blank control(Control)group.qPCR and Western blot assay were used to evaluate the TMEM97 silencing efficiency at mRNA and protein levels after siRNA-TMEM97 transfection.The proliferation ability of ovarian cancer cells was detected by CCK-8 assay after transfection.PI staining assay was used to observe the effect of down-regulated gene TMEM97 on cell cycle.The apoptosis rate was measured by Annexin V-FITC/PI assay.Western blot assay was used to evaluate the expressions of the B-cell lymphoma-2(Bcl-2),Bcl-2 associated X(Bax)and P38 mitogen-activated protein kinase pathway-associated proteins(P38/MAPK and p-P38/MAPK).Results Compared with the Control group and the siNC group,the proliferation ability of SKOV3 cells was inhibited in the si-TMEM97 group,and the early apoptosis rate of SKOV3 cells was significantly increased(P<0.01).The relative expression of Bcl-2 protein in SKOV3 cells was significantly decreased,and the relative expressions of Bax protein and p-P38/MAPK were significantly increased in si-TMEM97 group than those of Control group and siNC group(all P<0.05).Conclusion The down-regulation of TMEM97 decreases the proliferation and increases the apoptosis of SKOV3 cells,which may be related with the regulating the Bcl-2/Bax expression and activating P38/MAPK signaling pathway.

关 键 词：卵巢肿瘤 细胞增殖 细胞凋亡 BCL-2相关X蛋白质 P38丝裂原活化蛋白激酶类 TMEM97 P38/MAPK信号通路 

分 类 号：R737.31[医药卫生—肿瘤]