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小干扰RNA诱导的自噬相关基因7(Atg7)基因沉默对卵巢癌SKOV3细胞顺铂敏感性的影响

Effect of siRNA-induced Atg7 gene silencing on the sensitivity of ovarian cancer SKOV3 cells to cisplatin.

作者信息

Zhang Xuan, Wang Liang-Liang, Wang Beibei, Liu Hong-Li, Zhang Jing, Li Yan-Hua, Wang Li-Hua

机构信息

Department of Oncological Gynecology, First Affiliated Hospital of Bengbu Medical College Bengbu 233000, China.

出版信息

Am J Transl Res. 2020 May 15;12(5):2052-2061. eCollection 2020.

Abstract

Ovarian cancer is one of the most common types of gynecological malignant tumors. A proclivity for, or the development of chemoresistance severely affects treatment efficacy for ovarian cancer. Herein we found that as concentrations of cisplatin (DDP) used in SKOV3 cells increased, expression of intracellular reactive oxygen species (ROS) increased as did amounts of proteins of Beclin-1 and Autophagy-Related Gene 7 (Atg7) whereas in contrast, expression of P62 protein decreased gradually. Expression of Atg7 protein in SKOV3 cells in the siRNA-Atg7 transfection treatment group was significantly reduced compared to the negative control group. Post-application of DDP treatments, the apoptotic ratio of SKOV3 cells in the siRNA-Atg7 transfection group increased, and the cell survival rate decreased to a level significantly lower than in the negative control group. Cellular morphological analyses revealed remarkably decreased measures of cell density, as well as shrunk, deformed, and rounded cells with unclear boundaries, and revealed a decreased measures of mitochondrial membrane potential. Taken together, autophagy may be involved in the dynamics and mechanistics underlying DDP resistance in ovarian cancer SKOV3 cells. Thus, inhibition of autophagy through down-regulation expression of Atg7 may have beneficially enhanced the sensitivity of SKOV3 cells to DDP-based chemotherapy which could help improve treatment outcomes for patients afflicted by ovarian cancer.

摘要

卵巢癌是最常见的妇科恶性肿瘤类型之一。对化疗产生耐药性的倾向或耐药性的发展严重影响卵巢癌的治疗效果。在此我们发现,随着SKOV3细胞中顺铂(DDP)使用浓度的增加,细胞内活性氧(ROS)的表达增加,Beclin-1和自噬相关基因7(Atg7)的蛋白量也增加,而相比之下,P62蛋白的表达逐渐降低。与阴性对照组相比,siRNA-Atg7转染治疗组SKOV3细胞中Atg7蛋白的表达显著降低。应用DDP处理后,siRNA-Atg7转染组SKOV3细胞的凋亡率增加,细胞存活率降低至显著低于阴性对照组的水平。细胞形态学分析显示细胞密度显著降低,细胞出现萎缩、变形和边界不清的圆形,线粒体膜电位降低。综上所述,自噬可能参与卵巢癌SKOV3细胞对DDP耐药的机制和动态变化。因此,通过下调Atg7的表达来抑制自噬可能有助于增强SKOV3细胞对基于DDP的化疗的敏感性,从而有助于改善卵巢癌患者的治疗效果。

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