miR-30a-5p和NUAK1在非小细胞肺癌中的表达及对A549细胞增殖、迁移和侵袭的影响

Effects of miR-30a-5p on proliferation, migration and invasion of non-small cell lung cancer cells by directly targeting NUAK1

目的研究miR-30a-5p和靶基因NUAK1在非小细胞肺癌中的表达及对A549细胞增殖、迁移侵袭的影响。方法收集2016-09至2019-03在保定市第一医院行手术切除治疗的非小细胞肺癌患者癌组织和癌旁组织标本64例,QPCR和Western blot分别检测非小细胞肺癌组织样本和细胞中miR-30a-5p和NUAK1的表达;TargetScan网站预测miR-30a-5p与NUAK1间的靶向关系,双荧光素酶报告基因实验和Western blot进行验证;MTT和Transwell实验检测miR-30a-5p及联合过表达NUAK1对A549细胞增殖、迁移和侵袭的影响。结果非小细胞肺癌组织中miR-30a-5p的相对表达量(0.33±0.14)低于癌旁组织(1.00±0.21),差异有统计学意义(t=21.237,P<0.05);非小细胞肺癌组织中NUAK1 mRNA表达(4.13±1.87)和蛋白表达(3.41±1.62)均高于癌旁组织(1.00±0.17、1.00±0.16),差异有统计学意义(t=13.335、11.844,P<0.05)。非小细胞肺癌细胞系H460、H1299、A549中miR-30a-5p表达量(0.35±0.03、0.51±0.05、0.28±0.03)低于人正常肺上皮细胞BEAS-2B(1.00±0.11),差异有统计学意义(F=77.100,P<0.05);H460、H1299、A549细胞中NUAK1 mRNA相对表达量(2.98±0.30、2.39±0.24、3.42±0.36)和蛋白相对表达量(3.06±0.31、2.27±0.23、4.12±0.41)均显著高于BEAS-2B细胞(1.00±0.09、1.00±0.11),差异有统计学意义(F=46.660、63.070,P<0.05)。结论miR-30a-5p可通过靶向NUAK1抑制非小细胞肺癌细胞A549增殖、迁移和侵袭,提示miR-30a-5p和NUAK1可能成为一种新的临床诊断和治疗非小细胞肺癌的靶点。

Objective To investigate the expressions of miR-30 a-5 p and its target gene NUAK1 in non-small cell lung cancer,and the effects on the proliferation,migration and invasion of A549 cells.Methods The specimens of 64 cases of NSCLC were collected.QPCR and Western blot were used to detect the expressions of miR-30 a-5 p and NUAK1 in NSCLC tissues and cell lines.The relationship between miR-30 a-5 p and NUAK1 was predicted via TargetScan website and was verified by dual luciferase reporter assay and Western blot.The effects of miR-30 a-5 p on proliferation,migration and invasion of A549 were determined by MTT and Transwell assays.Results The expression of miR-30 a-5 p in NSCLC tissues(0.33±0.14)was significantly lower than in adjacent tissues(1.00±0.21)(t=21.237,P<0.05),while the expressions of NUAK1 mRNA(4.13±1.87)and protein(3.41±1.62)in NSCLC tissues were significantly higher than in adjacent tissues(1.00±0.17,1.00±0.16)(t=13.335,11.844,P<0.05).The expression of miR-30 a-5 p in NSCLC cells H460,H1299 and A549(0.35±0.03,0.51±0.05,0.28±0.03)was significantly lower than in normal human lung epithelial cells BEAS-2 B(1.00±0.08)(F=77.100,P<0.05).The expressions of NUAK1 mRNA(2.98±0.30,2.39±0.24,3.42±0.35)and protein(3.06±0.31,2.27±0.23,4.12±0.41)in H460,H1299 and A549 cells were significantly higher than in BEAS-2 B cells(1.00±0.09,1.00±0.11)(F=46.660,63.070,P<0.05).Conclusions miR-30 a-5 p can inhibit the proliferation,migration and invasion of A549 cells by targeting NUAK1,suggesting that miR-30 a-5 p and NUAK1 may become new targets for diagnosis and treatment of NSCLC.