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

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（cleavedcas3）/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。

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

Objective To investigate the effect of interfering with long non-coding RNA（ lncRNA） MNX1-AS1 onthe 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 oflncRNA MNX1-AS1， and the sequence with the most obvious interference effect was screened ； the spheroidizationof stem cells was observed by microscopy ； the expression levels of stem cell marker protein OCT4， SOX2， andABCG2 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 commendedto test changes in mitochondrial membrane potential ； Western blotting was performed to measure the expression ofmitochondrial damage marker proteins Bax/Bcl-2 ， cleaved cas3/cas3 and c-Myc. Results The comparison of the threesequences showed that the interference in the MNX1-AS1-shRNA3 group was the most significant. This group wasselected as the interference group for the follow-up experiment and divided into the control group， negative controlgroup（ shRNA-NC） and interference group（ MNX1-AS1-shRNA3）. Compared with the control group， the numberand diameter of SKOV3 stem cells in the MNX1-AS1-shRNA3 interference group were reduced， and the expressionlevels of stem cell marker proteins OCT4， SOX2， ABCG2 were significantly reduced. SOD activity and c-Mycprotein expression were both significantly decreased， the expression of mitochondrial damage marker proteins（ Bax/Bcl-2， cleaved cas3/cas3）， MDA， and ROS contentwere significantly increased，the membrane potential ofmitochondria decreased. Conclusion Interfering withthe lncRNA MNX1-AS1 induces the decrease of stemlike properties， the enhancment of oxidative stress， and the aggravation of mitochondrial damage in ovarian cancercells. Interfering with the lncRNA MNX1-AS1 has a significant inhibitory effect on SKOV3 cells and has the potentialof targeted treatment of ovarian cancer.