干扰长链非编码RNA MNX1-AS1对卵巢癌细胞的干样特性、氧化应激以及线粒体功能损伤的影响  

Effect of interfering with long non-coding RNA MNX1-AS1 on the stem-like characteristics,oxidative stress and mitochondrial function damage of ovarian cancer cells

作　　者：张晓丹[1] 徐毅[1] 杜德奇[1] 郭莹[2] 王鲁文 Zhang Xiaodan;Xu Yi;Du Deqi;Guo Ying;Wang Luwen(Department of Obstetrics,Zhengzhou Maternal and Child Health Care Hospital,Zhengzhou 450000;Department of Gynecology,Zhengzhou Maternal and Child Health Care Hospital,Zhengzhou 450000;Department of Gynecology,The Third Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]郑州市妇幼保健院产科,郑州450000 [2]郑州市妇幼保健院妇科,郑州450000 [3]郑州大学第三附属医院妇科,郑州450000

出　　处：《解剖学杂志》2021年第5期394-398,共5页Chinese Journal of Anatomy

基　　金：河南省科技攻关计划项目(201803243)。

摘　　要：目的:探讨干扰长链非编码RNA(lncRNA)MNX1-AS1对卵巢癌细胞的干样特性、氧化应激以及线粒体功能损伤的影响。方法:将KOV3细胞随机分组,进行处理及培养,RT-PCR检测RNA MNX1-AS1的表达,筛选干扰效果最明显的序列;观察干细胞成球情况;免疫印迹检测干细胞标记物蛋白八聚体结合转录因子4(OCT4)、性别决定区Y框蛋白(SOX2)和ATP结合转运蛋白G超家族成员2(ABCG2)的表达;检测氧化应激标记物丙二醛(MDA)、超氧化物歧化酶(SOD)的含量;荧光探针检测活性氧(ROS)的含量;流式细胞仪检测线粒体膜电位的变化;免疫印迹检测B淋巴细胞瘤-2基因(Bcl-2)/Bcl-2相关X蛋白(Bax)、活化胱天蛋白酶3(cleaved cas3)/cas3、c-Myc的表达。结果:3个序列对比显示MNX1-AS1-shRNA3干扰组最为显著,选择此组为后续实验干扰组,分组为对照组、阴性对照组(shRNA-NC)、干扰组(MNX1-AS1-shRNA3);与对照组相比,MNX1-AS1-shRNA3干扰组SKOV3干细胞成球数量、直径减小,干细胞标记物蛋白OCT4、SOX2、ABCG2的表达显著降低,SOD活性、c-Myc蛋白表达均显著降低,线粒体损伤标记物蛋白的表达(Bax/Bcl-2、cleaved cas3/cas3)、MDA与ROS含量均显著升高,线粒体的膜电位下降。结论:干扰lncRNA MNX1-AS1诱导卵巢癌细胞的干样特性降低、氧化应激增强、线粒体损伤加重,干扰lncRNA MNX1-AS1对SKOV3细胞抑制效果显著,具有靶向治疗卵巢癌的潜力。Objective:To investigate the effect of interfering with long non-coding RNA(lncRNA)MNX1-AS1 on the stem-like characteristics,oxidative stress and mitochondrial function damage of ovarian cancer cells.Methods:SKOV3 cells were randomly grouped,processed and cultured.RT-PCR was used to detect the expression level of lncRNA MNX1-AS1,and the sequence with the most obvious interference effect was screened;the spheroidization of stem cells was observed by microscopy;the expression levels of stem cell marker protein OCT4,SOX2,and ABCG2 were detected by Western blotting;the content of oxidative stress markers(MDA,SOD)was detected;fluorescent probe was used to detect the content of reactive oxygen species(ROS);flow cytometry was commended to test changes in mitochondrial membrane potential;Western blotting was performed to measure the expression of mitochondrial damage marker proteins Bax/Bcl-2,cleaved cas3/cas3 and c-Myc.Results:The comparison of the three sequences showed that the interference in the MNX1-AS1-shRNA3 group was the most significant.This group was selected as the interference group for the follow-up experiment and divided into the control group,negative control group(shRNA-NC)and interference group(MNX1-AS1-shRNA3).Compared with the control group,the number and diameter of SKOV3 stem cells in the MNX1-AS1-shRNA3 interference group were reduced,and the expression levels of stem cell marker proteins OCT4,SOX2,ABCG2 were significantly reduced.SOD activity and c-Myc protein expression were both significantly decreased,the expression of mitochondrial damage marker proteins(Bax/Bcl-2,cleaved cas3/cas3),MDA,and ROS content were significantly increased,the membrane potential of mitochondria decreased.Conclusion:Interfering with the lncRNA MNX1-AS1 induces the decrease of stem-like properties,the enhancment of oxidative stress,and the aggravation of mitochondrial damage in ovarian cancer cells.Interfering with the lncRNA MNX1-AS1 has a significant inhibitory effect on SKOV3 cells and has the potential of targeted tr

关 键 词：卵巢癌 lncRNA MNX1-AS1 SKOV3细胞 干细胞特性 氧化应激 线粒体 

分 类 号：R737.31[医药卫生—肿瘤]