miRNA-373在MDA-MB-231乳腺癌细胞中的表达及对生物学行为的影响研究

Expression of miRNA-373 in MDA-MB-231 breast cancer cells and its effect on biological behavior

目的探讨miRNA-373对乳腺癌细胞MDA-MB-231增殖、侵袭等生物学行为的影响及其机制。方法采用定量聚合酶链反应(qPCR)检测MDA-MB-231、MDA-MB-435、MCF-7、ZR-75-1乳腺癌细胞株中miRNA-373的表达情况;MDA-MB-231细胞株中分别转染miRNA-373(NC组)和miRNA-373mimics(miRNA-373mimics组),采用CCK-8实验和Transwell实验检测乳腺癌细胞增殖和侵袭能力,细胞划痕实验检测细胞迁移情况,流式细胞仪检测细胞凋亡情况,Westernblot法检测蛋白激酶B(AKT)、磷酸化蛋白激酶B(p-AKT)的表达情况。结果MDA-MB-231乳腺癌细胞中miRNA-373的表达量相对MDA-MB-435、MCF-7、ZR-75-1细胞较低。miRNA-373mimics组乳腺癌细胞中miRNA-373、p-AKT表达量均高于NC组细胞,差异均有统计学意义(P﹤0.05)。CCK8实验检测结果显示,细胞在转染24、48、72h时,NC组和miRNA-373mimics组细胞的吸光度值比较,差异均无统计学意义(P﹥0.05)。流式细胞仪检测NC组和miRNA-373mimics组细胞的凋亡情况,两组细胞凋亡率比较,差异无统计学意义(P﹥0.05)。转染24、48h时,NC组细胞迁移距离分别短于miRNA-373mimics组,差异均有统计学意义(P﹤0.05)。结论miRNA-373可诱导乳腺癌细胞侵袭及迁移,其机制可能是通过增强AKT信号通路的磷酸化水平来促进乳腺癌细胞的迁移和侵袭。

Objective To investigate the effect of miRNA-373 on biological behavior like proliferation and invasion of breast cancer cell MDA-MB-231 and its mechanism. Method Quantitative polymerase chain reaction (qPCR) was used to detect the expression of miRNA-373 in breast cancer cell lines MDA-MB-231, MDA-MB-435, MCF-7 and ZR- 75-1. miRNA-373 (NC group) and miRNA-373 mimics (miRNA-373 mimics group) were transfected into MDA-MB- 231 cell line respectively, and the proliferation and invasion of breast cancer cells were detected by CCK-8 assay and Transwell assay. Cell apoptosis was detected by flow cytometry assay and cell migration was detected by cell scratch assay. The expression of protein kinase B (AKT) and phosphorylated protein kinase B (p-AKT) were detected by Western blot. Result The expression of miRNA-373 in breast cancer cell MDA-MB-231 was lower than that in MDA-MB-435, MCF-7, ZR-75-1 cells and the expression level of miRNA-373, p-AKT in miRNA-373 mimics group was higher than those in NC group, the differences were statistically significant (P<0.05). There was no significant difference in the optical density value between NC group and miRNA-373 mimics group at 24, 48, 72 h after transfection according to the CCK8 assay (P>0.05). Flow cytometry assay was used to detect the apoptosis of cells in NC group and miRNA-373 mimics group and the difference between the two groups was not statistically significant (P>0.05). The migration distance of NC group at 24, 48 h after transfection was shorter than that of miRNA-373 mimics group with statistically significant difference (P<0.05). Conclusion miRNA-373 can promote the invasion and migration of breast cancer cell, and the mechanism may be induced by the enhancement of the phosphorylation of AKT signaling pathway.