miRNA-409-3p /Beclin-1 信号轴在小细胞肺癌化疗耐药的作用及分子机制研究  

Molecular Mechanism of miRNA-409-3p /Beclin-1 Signal Axis in Chemotherapy Resistance in Small Cell Lung Cancer

作　　者：韩忠诚[1] 苏莹[1] 张玲玲[1] 柳江[1] 徐腾飞 Han Zhongcheng;Su Ying;Zhang Lingling;Liu Jiang;Xu Tengfei(People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830000,Xinjiang,China;Department of General Medicine,People’s Hospital of Pishan County,Xinjiang Uygur Autonomous Region,Hetian 848000,Xinjiang,China)

机构地区：[1]新疆维吾尔自治区人民医院肿瘤科,乌鲁木齐830000 [2]新疆维吾尔自治区人民医院皮山医院全科医疗科,新疆和田848000

出　　处：《肿瘤预防与治疗》2020年第9期729-738,共10页Journal of Cancer Control And Treatment

基　　金：新疆维吾尔自治区自然科学基金项目(编号:2019D01C100)。

摘　　要：目的: 探讨 miRNA-409-3p /Beclin-1 信号轴在小细胞肺癌耐药中的作用机制。方法: qRT-PCR 检测用于分析基因表达情况;Western blot 检测蛋白表达情况;mimic NC、miRNA-409-3p mimic、pcDNA3. 1 质粒和 pcDNA3. 1-Bec- lin-1 质粒分别转染依托泊苷( VP16) 和顺铂( DDP) 耐药的小细胞肺癌细胞系 SBC-5 /EP;荧光素酶活性分析实验用于检测基因靶向关系;GFP-LC3 绿色荧光融合蛋白实验用于检测细胞自噬;克隆形成实验和流式细胞术实验用于检测细胞的增殖和凋亡;MTT 法用于检测细胞对 VP16-DDP 的敏感性。结果: 在 VP16-DDP 耐药的小细胞肺癌组织和小细胞肺癌细胞( SBC-5 /EP) 中,miRNA-409-3p 表达量低,Beclin-1 表达量高;荧光素酶活性分析实验显示 miRNA-409- 3p 靶向作用于 Beclin-1;miRNA-409-3p mimic 可抑制 SBC-5 /EP 细胞自噬和增殖,促进细胞凋亡,提高 SBC-5 /EP 细胞对 VP16-DDP 的敏感性,抑制 PI3KC3 和 Vpsl5 蛋白表达。pcDNA3. 1-Beclin-1 可逆转 miRNA-409-3p mimic 对 SBC-5 / EP 细胞自噬和增殖的抑制作用、对细胞凋亡的促进作用、对 VP16-DDP 药物敏感性的提高作用,以及对 PI3KC3 和 Vpsl5 蛋白表达的抑制作用。结论: miRNA-409-3p 通过靶向作用于 Beclin-1 而抑制 VP16-DDP 耐药的小细胞肺癌细胞的自噬和增殖,促进细胞凋亡,提高耐药的小细胞肺癌细胞对 VP16-DDP 的药物敏感性,可能与 PI3KC3 /Vpsl5 信号通路相关。Objective:To investigate the molecular mechanism of miRNA-409-3p/Beclin-1 signal axis in chemoresistant small cell lung cancer.Methods:qRT-PCR was used to analyze gene expressions;western blot was used to detect protein expressions;mimic NC,miRNA-409-3p mimic,plas-mid pcDNA3.1 and plasmid pcDNA3.1-Beclin-1 were trans-fected into etoposide combined with cisplatin(VP16-DDP)- resistant small cell lung cancer cell line ( SBC-5 /EP);luciferase activity assay was used to detect the targeting relationship between genes;GFP-LC3 dot formation was used to detect cell autophagy;clone formation experiment and flow cytometry were used to detect cell proliferation and apoptosis;MTT was used to detect the sensitivity of VP16-DDP. Results: In VP16- DDP-resistant small cell lung cancer tissues and cell line ( SBC-5 /EP) ,the expression level of miRNA-409-3p was down- regulated,while that of Beclin-1 was up-regulated. Luciferase activity assay showed that miRNA-409-3p targeted Beclin-1. miRNA-409-3p mimic inhibited autophagy and proliferation,promoted apoptosis,improved the sensitivity of VP16-DDP and inhibited the expression of PI3KC3 and Vpsl5 proteins in the SBC-5 /EP cell. miRNA-409-3p mimic inhibits cell autophagy and proliferation,promotes cell apoptosis,increases the sensitivity to VP16-DDP and inhibits the expressions of PI3KC3 and Vpsl5;and the foregoing could be reversed by pcDNA3. 1-Beclin-1. Conclusion: miRNA-409-3p negatively regulates Bec- lin-1 to inhibit the autophagy and proliferation of VP16-DDP-resistant small cell lung cancer cells,promotes apoptosis,and improves the sensitivity of VP16-DDP-resistant small cell lung cancer cells,which may involve PI3KC3 /Vpsl5 signaling pathway.

关 键 词：miRNA-409-3p BECLIN-1 细胞自噬 VP16-DDP耐药 PI3KC3/Vpsl5信号通路 小细胞肺癌 细胞增殖 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤] Q74[医药卫生—临床医学]