| 摘要: |
| 目的 探究miRNA-4286在非小细胞肺癌细胞对顺铂敏感性中的调节作用及相关机制。方法 采用qPCR法检测人非小细胞肺癌A549细胞和顺铂耐药A549细胞（A549/DDP细胞）中miRNA-4286的表达水平。采用miRNA-4286模拟物和抑制剂构建miRNA-4286的过表达和低表达细胞模型，分为过表达阴性对照组、过表达miRNA-4286组、低表达阴性对照组、低表达miRNA-4286组。采用CCK-8法、平板集落形成实验和流式细胞术检测miRNA-4286表达对非小细胞肺癌细胞的顺铂敏感性影响；采用TargetScan、qPCR法和蛋白质印迹法检测miRNA-4286和核输出蛋白1（XPO1）的靶向关系；采用蛋白质印迹法检测在顺铂的作用下改变miRNA-4286后非小细胞肺癌细胞的凋亡蛋白胱天蛋白酶3（caspase 3）、B淋巴细胞瘤2相关X蛋白（Bax）表达量的改变。结果 miRNA-4286在A549细胞中的表达高于A549/DDP细胞（P＜0.01）。在0～2 μg/mL顺铂的作用下，过表达miRNA-4286后A549细胞增殖率降低、凋亡率增高（P＜0.05或P＜0.01） ；低表达miRNA-4286后结果与之相反（P＜0.05或P＜0.01）。过表达miRNA-4286后A549细胞中XPO1的mRNA和蛋白表达均降低（P＜0.05或P＜0.01），低表达miRNA-4286后结果与之相反（P＜0.05或P＜0.01）。在顺铂作用下，下调miRNA-4286抑制A549细胞caspase 3、Bax的表达（P＜0.05或P＜0.01），而下调XPO1能负调控这一作用（P＜0.05或P＜0.01）。结论 miRNA-4286通过靶向XPO1调控非小细胞肺癌A549细胞对顺铂的敏感性，其机制可能与细胞的凋亡通路相关。 |
| 关键词: 微RNA-4286 非小细胞肺癌 顺铂 肿瘤耐药性 细胞凋亡 |
| DOI：10.16781/j.CN31-2187/R.20211045 |
| 投稿时间：2021-10-19修订日期：2022-03-07 |
| 基金项目: |
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| MicroRNA-4286 regulates the sensitivity of non-small cell lung cancer cells to cisplatin by targeting exportin 1 |
| WANG Pei-rong,JIANG Tao* |
(Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the regulatory effect and related mechanism of microRNA (miRNA)-4286 on sensitivity of non-small cell lung cancer cells to cisplatin. Methods The expression of miRNA-4286 in non-small cell lung cancer A549 cells and A549 cisplatin resistant cells (A549/DDP) was detected by quantitative polymerase chain reaction (qPCR). The overexpression and low expression cell models of miRNA-4286 were successfully constructed by using miRNA-4286 mimics and inhibitor, respectively, and were divided into overexpression negative control group, overexpression miRNA-4286 group, low expression negative control group, and low expression miRNA-4286 group. Cell counting kit 8 (CCK-8), plate colony-forming test and flow cytometry were used to detect the effect of miRNA-4286 expression on the sensitivity of non-small cell lung cancer cells to cisplatin. The targeting relationship between miRNA-4286 and exportin 1 (XPO1) was detected by TargetScan, qPCR, and Western blotting. Western blotting was used to detect the expression of apoptotic proteins cysteine aspartic acid specific protease 3 (caspase 3), B-cell lymphoma-related X protein (Bax) in non-small cell lung cancer cells after changing miRNA-4286 under the action of cisplatin. Results The expression of miRNA-4286 in A549 cells was significantly higher than that in A549/DDP cells (P<0.01). Under the action of 0-2 μg/mL cisplatin, the proliferation rate of A549 cells was decreased and the apoptosis rate was increased after overexpression of miRNA-4286 (P<0.05 or P<0.01); the results were opposite with low expression of miRNA-4286 (P<0.05 or P<0.01). The mRNA and protein expression levels of XPO1 in A549 cells were decreased after overexpression of miRNA-4286 (P<0.05 or P<0.01), the results were opposite with low expression of miRNA-4286 (P<0.05 or P<0.01). Under the action of cisplatin, down-regulation of miRNA-4286 inhibited the expression of apoptotic proteins caspase 3 and Bax in A549 cells, and down-regulation of XPO1 could negatively regulate this effect (P<0.05 or P<0.01). Conclusion miRNA-4286 can target XPO1 and regulate the sensitivity of non-small cell lung cancer A549 cells to cisplatin, and the mechanism may be related to the apoptosis pathway. |
| Key words: microRNA-4286 non-small cell lung cancer cisplatin tumor drug-resistance apoptosis |
