miR-30a-5p对肺癌细胞增殖与迁移的影响及其机制

miR-30a-5p inhibits proliferation and migration of non-small cell lung cancer cell and its mechanism

摘要:

研究miR-30a-5p 对非小细胞肺癌（NSCLC）增殖和迁移的影响，并探讨其调控机制。方法NSCLC A549细胞系分成两组，miR-30a-5p组和对照组，分别转染miR-30a-5p mimics和microRNA scramble，MTT实验及细胞划痕实验分别测定两组细胞增殖和迁移能力，实时聚合酶链反应（real time-PCR）和蛋白免疫印迹法（Western blot）分别测定两组泛素蛋白连接酶（UBE3C）mRNA 和蛋白质表达水平。结果培养48 h后，miR-30a-5p 组相对活细胞百分比低于对照组[（203.7±16.8）% vs（330.4±20.7）%（p =0.000）]；培养72 h后，miR-30a-5p组相对活细胞百分比（258±30.2）%，对照组相对活细胞百分比（403.4±28.4）%，miR-30a-5p组相对活细胞百分比低于对照组（p =0.001）。细胞划痕实验示：miR-30a-5p组划痕愈合率为（23.2±2.7）%，对照组划痕愈合率为（89.2±4.8）%，miR-30a-5p 组划痕愈合率低于对照组（p =0.000）。real time-PCR显示，miR-30a-5p 组UBE3C mRNA 表达水平为（0.15±0.02），对照组UBE3C mRNA 表达水平为（1.03±0.09），miR-30a-5p组UBE3C mRNA表达水平低于对照组（p =0.000）；Western blot显示，miR-30a-5p组UBE3C 蛋白表达水平为（1.57±0.26），对照组UBE3C 蛋白表达水平为（5.32±0.42），miR-30a-5p 组UBE3C 蛋白表达水平低于对照组（p =0.000）。结论miR-30a-5p抑制NSCLC 增殖和迁移，其机制与下调UBE3C表达有关。

Abstract:

To investigate the role of miR-30a-5p on the regulation of the proliferation and migration of NSCLC, and its regulatory mechanism. Methods A549 cells were divided into two groups, miR-30a-5p group (transfected with miR-30a-5p mimics) and control group (transfected with microRNA scrambles).The proliferation and migration ability was measured by MTT assay and wound healing experiment, respectively.The expression level of UBE3C mRNA and protein was measured by RT-PCR and Western blot. Results The relative percentage of living cells in miR-30a-5p group was significantly lower than control group after cultur-ing for 48 h, (203.7 ±16.8)% vs (330.4 ±20.7)%, p= 0.000. There was (258 ±30.2)% of relative percentage of living cells in miR-30a-5p group, which was significantly lower than (403.4 ±28.4)% in the control group (p =0.001). The wound healing rate was (23.2 ±2.7%) in miR-30a-5p group, which was significantly lower than(89.2 ±4.8)% in the control group (p = 0.000). RT-PCR indicated that the expression level of UBE3C mRNAwas (0.15 ±0.02) in miR-30a-5p group, which was significantly lower than (1.03 ±0.09) in the control group( p= 0.000). The expression level of UBE3C protein was (1.57 ±0.26) in miR-30a-5p group, which was signifi-cantly lower than (5.32 ±0.42) in control group ( p= 0.000). Conclusions MiR-30a-5p inhibits cell prolifera-tion and migration of non-small cell lung cancer cell by suppressing UBE3C gene expression.