miR-338-5p过表达对胃癌细胞进展及耐药基因表达的影响

Effect of miR-338-5p overexpression on progression of gastric cancer cells and drug-resistant gene expression

目的探讨miR-338-5p过表达对胃癌细胞侵袭和耐药性的影响。方法人胃癌细胞MKN-45分为mimics组、mimics-NC组、inhibitor组和inhibitor-NC组,分别转染miR-338-5p-mimics、miR-338-5p-inhibitor及其NC质粒,未行转染的人胃癌细胞MKN-45为对照组。在转染培育24 h后,用CCK-8检测细胞的增殖活性,体外侵袭实验测定细胞的迁移及侵袭能力;使用qRT-PCR检测各组细胞中的耐药基因P-糖蛋白(PGP)、三磷酸腺苷结合盒转运蛋白B2(ATP-binding cassette transporter B2,ABCB2)和G2(ABCG2)相关mRNA的表达;Western blot测定各组细胞中的第10染色体同源丢失性磷酸酶张力蛋白基因(phosphatase and tensin homology deleted on chromosome ten,PTEN)、蛋白激酶B(protein kinase B,AKT)和磷酸化蛋白激酶B(phosphoprotein kinases B,p-AKT)蛋白表达。结果与对照组和mimics-NC组相比,mimics组细胞增殖活力下降(P<0.05),细胞的迁移侵袭能力、耐药性相关PGP、ABCB2、ABCG2的mRNA表达下降(P<0.05);细胞中PTEN表达量上升(P<0.05),p-AKT蛋白表达量下降(P<0.05),AKT蛋白无显著变化(P>0.05)。与对照组和inhibitor-NC组相比,inhibitor组细胞抑制率下降(P<0.05),细胞迁移侵袭能力、PGP、ABCB2、ABCG2的mRNA表达上升(P<0.05),PTEN表达量下降(P<0.05),p-AKT蛋白表达量上升(P<0.05),AKT蛋白无显著变化(P>0.05)。结论miR-338-5p影响PTEN/AKT通路的表达,抑制耐药基因表达和胃癌细胞的侵袭迁移,从而抑制胃癌细胞的增殖。

Objective To investigate the effect of miR-338-5p overexpression on the invasion and the drug resistance of gastric cancer cells.Methods Human gastric cancer cells MKN-45 were divided into mimics group,mimics-NC group,inhibitor group and inhibitor-NC group,and transfected with miR-338-5p-mimics,miR-338-5p-inhibitor and its corresponding NC plasmid respectively.Human gastric cancer cells MKN-45 without transfection were chosen as control group.At 24 h after transfection,the proliferation activity of the cells was measured by CCK-8,and the migration and invasion abilities of the cells were determined by in vitro invasion assay.RT-qPCR was used to detect the mRNA expression of the drug resistance genes P-glycoprotein(PGP),adenosine triphosphate-binding cassette transporter B2(ABCB2),and G2(ABCG2)in each group.The expression of phosphatase and tensin homolog deleted on chromosome ten(PTEN),protein kinase B(AKT),and phosphoprotein kinases B(p-AKT)was determined by Western blot.Results Compared with control group and mimics-NC group,the proliferation viability of cells was decreased in mimics group(P<0.05),the expression of migratory and invasive abilities and the drug resistance-related mRNA(PGP,ABCB2,ABCG2)levels were decreased(P<0.05),the expression of PTEN in cells was increased(P<0.05),and the expression of p-AKT protein was decreased(P<0.05),while there was no significant change in AKT protein expression(P>0.05).Compared with control group and inhibitor-NC group,the cell inhibition was decreased in inhibitor group(P<0.05),the cell migration and invasion abilities and the mRNA expression of PGP,ABCB2 and ABCG2 were increased(P<0.05),PTEN expression was decreased(P<0.05),and p-AKT protein expression was increased(P<0.05),while there was no significant change in AKT protein(P>0.05).Conclusion MiR-338-5p affects the expression of PTEN/AKT pathway,and inhibits the drug-resistant gene expression and the invasion and migration of gastric cancer cells,thus suppressing the proliferation of gastric cancer cells.