miR-152-3p对非小细胞肺癌A549细胞顺铂敏感性的作用及分子机制研究

Effect of miR-152-3p on cisplatin sensitivity in non-small cell lung cancer A549 cells and its molecular mechanism

目的:探究miR-152-3p对非小细胞肺癌A549细胞顺铂敏感性的作用及其分子机制。方法:采用顺铂处理体外培养的A549细胞,检测细胞内miR-152-3p、SOS1表达水平的变化;采用荧光素酶活性检测法分析miR-152-3p与SOS1的调控关系。A549细胞转染miR-152-3p慢病毒过表达载体、SOS1慢病毒过表达载体或慢病毒空载体,采用顺铂处理转染的细胞,比较miR-152-3p过表达或SOS1过表达对A549细胞增殖、细胞周期、凋亡及迁移的影响。结果:与正常培养的A549细胞相比,顺铂处理的A549细胞内miR-152-3p表达水平显著降低,而SOS1表达水平显著升高(P<0.05);生物信息学分析显示SOS1是miR-152-3p的一个潜在靶基因,而荧光素酶活性检测法分析显示miR-152-3p具有负调控SOS1的作用。与转染空载体的A549细胞相比,miR-152-3p过表达显著抑制A549细胞增殖及迁移、阻滞细胞周期、促进细胞凋亡(P<0.05)。miR-152-3p过表达的A549细胞内SOS1表达水平显著低于转染空载体的细胞(P<0.05);与单独转染miR-152-3p过表达载体的细胞相比,同时转染miR-152-3p过表达载体和SOS1慢病毒过表达载体的A549细胞增殖、迁移增强,且细胞凋亡受到显著抑制(P<0.05)。结论:miR-152-3p通过抑制SOS1表达增强A549细胞对顺铂的敏感性,继而抑制A549细胞增殖、迁移,阻滞细胞周期以及促进A549细胞凋亡。

Objective:To explore the effect and molecular mechanism of miR-152-3p on cisplatin sensitivity of non-small cell lung cancer A549 cells.Methods:A549 cells cultured in vitro were treated with cisplatin to detect the changes of miR-152-3p and SOS1 expression levels.The regulatory relationship between miR-152-3p and SOS1 was analyzed by luciferase activity assay.A549 cells were transfected with miR-152-3p,SOS1 lentivirus overexpression vector or empty lentivirus vector,and then treated with cisplatin to compare the effects of miR-152-3p or SOS1 overexpression on the proliferation,cell cycle,apoptosis and migration of A549 cells.Results:Compared with normal cultured A549 cells,the expression level of miR-152-3p in A549 cells treated with cisplatin was significantly decreased,while the expression level of SOS1 was significantly increased(P<0.05).Bioinformatics analysis showed that SOS1 was a potential target gene of miR-152-3p,while luciferase activity assay showed that miR-152-3p had a negative regulation effect on SOS1.Compared with A549 cells transfected with empty vector,miR-152-3p overexpression inhibited proliferation and migration,blocked cell cycle and promoted apoptosis significantly(P<0.05).The expression level of SOS1 in A549 cells transfected with miR-152-3p overexpression vector was significantly lower than that of cells transfected with empty vector(P<0.05).The expression level of SOS1 in A549 cells transfected with miR-152-3p overexpression vector was significantly lower than that of cells transfected with empty vector(P<0.05).Compared with single transfection miR-152-3p expression vector of the cell,the proliferation and migration of A549 cell transfected with miR-152-3p expression vector were enhanced,and apoptosis was suppressed(P<0.05).Conclusion:miR-152-3p can enhance the sensitivity of A549 cells to cisplatin by inhibiting the expression of SOS1,and then inhibit A549 cells proliferation and migration,block cell cycle and promote A549 cell apoptosis.