下调lncRNA LINC00176对肺癌A549/DDP细胞顺铂耐药和自噬的影响  

Effects of down-regulating lncRNA LINC00176 on cisplatin resistance and autophagy in lung cancer A549/DDP cells

作　　者：崔华[1] 燕红霞 尹梅[1] CUI Hua;YAN Hongxia;YIN Mei(Department of Respiratory and Critical Medicine,General Hospital of Ningxia Medical University,Yinchuan 750001,Ningxia,China;Department of Neurology,the Second People's Hospital of Yinchuan 750011,Ningxia,China)

机构地区：[1]宁夏医科大学总医院呼吸与危重症医学科,宁夏银川750001 [2]银川市第二人民医院神经内科,宁夏银川750011

出　　处：《中国临床药理学与治疗学》2021年第6期616-623,共8页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：宁夏回族自治区自然科学基金项目(2017A0484)。

摘　　要：目的:研究下调lncRNA LINC00176对肺癌A549/DDP细胞顺铂耐药和自噬的影响及机制。方法:qRT-PCR方法测定正常支气管上皮16HBE细胞和肺癌A549、A549/DDP、NCI-H1299、SK-MES-1细胞中LINC00176的表达。将A549/DDP细胞分成对照组、si-NC组、si-LINC00176组、si-LINC00176+Anti-miR-NC组、si-LINC00176+Anti-miR-138-5p组。MTT实验检测顺铂对A549/DDP的半数抑制浓度(IC50)。流式细胞术测定细胞凋亡。Western blot测定LC3-I、LC3-II、Beclin 1、C-Caspase-3蛋白表达。利用荧光素酶报告系统鉴定LINC00176和miR-138-5p的靶向关系。结果:与16HBE细胞比较,A549、A549/DDP、NCI-H1299、SK-MES-1细胞中LINC00176表达水平显著升高(P<0.01)。与A549细胞比较,A549/DDP细胞中LINC00176表达水平显著升高(P<0.01)。与对照组、si-NC组比较,si-LINC00176组A549/DDP细胞IC50显著降低(P<0.01),LC3-II/LC3-I、Beclin 1蛋白表达显著降低(P<0.01),凋亡率、C-Caspase-3蛋白表达、miR-138-5p表达水平显著升高(P<0.01)。LINC00176与miR-138-5p直接结合。与si-LINC00176+Anti-miR-NC组比较,si-LINC00176+Anti-miR-138-5p组A549/DDP细胞IC50显著升高(P<0.01),LC3-II/LC3-I、Beclin 1蛋白表达显著升高(P<0.01),凋亡率、C-Caspase-3蛋白表达显著降低(P<0.01)。结论:下调lncRNA LINC00176通过靶向上调miR-138-5p能够抑制肺癌A549/DDP细胞顺铂耐药,抑制细胞自噬,诱导细胞凋亡。AIM:To study the effects and mechanism of down-regulating lncRNA LINC00176 on cisplatin resistance and autophagy in lung cancer A549/DDP cells.METHODS:The qRT-PCR method was used to determine the expression changes of LINC00176 in normal bronchial epithelial 16HBE cells and lung cancer A549,A549/DDP,NCI-H1299 and SK-MES-1 cells.A549/DDP cells were divided into Control group,si-NC group,and si-LINC00176 group,si-LINC00176+Anti-miR-NC group,and si-LINC00176+Anti-miR-138-5p group.MTT experiment detected the half inhibitory concentration(IC 50)of cisplatin on A549/DDP.Flow cytometry was used to measure cell apoptosis.Western blot was used to determine the protein expression of LC3-I,LC3-II,Beclin 1,C-Caspase-3.The luciferase reporter system was used to identify the targeting relationship between LINC00176 and miR-138-5p.RESULTS:The expression level of LINC00176 in A549,A549/DDP,NCI-H1299 and SK-MES-1 cells was higher than that in 16HBE cells(P<0.01).The expression level of LINC00176 in A549/DDP cells was significantly higher than that in A549 cells(P<0.01).Compared with the Control and si-NC groups,the IC 50 of A549/DDP cells in the si-LINC00176 group was significantly reduced(P<0.01),the expression of LC3-II/LC3-I,Beclin 1 protein was significantly reduced(P<0.01),the apoptosis rate,and the expression of C-Caspase-3 protein and miR-138-5p were significantly increased(P<0.01).LINC00176 directly bound to miR-138-5p.Compared with the si-LINC00176+Anti-miR-NC group,the IC 50 of A549/DDP cells in the si-LINC00176+Anti-miR-138-5p group was significantly increased(P<0.01),and the expression of LC3-II/LC3-I and Beclin 1 protein was significantly increased(P<0.01),the apoptosis rate and C-Caspase-3 protein expression were significantly reduced(P<0.01).CONCLUSION:Down-regulating lncRNA LINC00176 could inhibit cisplatin resistance,inhibit autophagy and induce apoptosis in lung cancer A549/DDP cells by targeting and up-regulating miR-138-5p.

关 键 词：肺癌 顺铂耐药 自噬 LINC00176 miR-138-5p 

分 类 号：R965.2[医药卫生—药理学] R734.2[医药卫生—药学]