miR-96在氯菊酯促MCF-7细胞迁移过程中的调控作用

Regulation of miR-96 in permethrin-induced migration of MCF-7 cells

[目的]miRNA参与氯菊酯致细胞的毒理机制还不够清楚。旨在研究miR-96在低剂量氯菊酯诱导细胞迁移中的调控作用,为氯菊酯毒理机制提供新的补充。[方法]通过向人乳腺细胞MCF-7中转染miR-96类似物和抑制剂,运用荧光定量PCR法检测miR-96的相对表达量,人为过表达和抑制细胞中的miR-96;通过细胞划痕愈合试验检测过表达和抑制miR-96后,氯菊酯对细胞迁移能力的影响。[结果]以上试验显示,细胞中miR-96的表达量跟氯菊酯的作用剂量呈正相关;100μmol/L氯菊酯作用48 h时,细胞中miR-96的表达量最高,为3.68±0.43,差异极显著(P<0.01)。向MCF-7细胞转染miR-96类似物和抑制剂后,miR-96表达分别上调2倍和下调4倍。继续对细胞施加100μmol/L氯菊酯,过表达miR-96可以增强氯菊酯促细胞迁移愈合能力;反之,抑制miR-96能降低氯菊酯的该项能力。[结论]研究证实miR-96参与了氯菊酯致MCF-7细胞的迁移作用,氯菊酯在影响细胞生理活动过程中受miRNA的调控。

[Aims] The toxicological mechanism of miRNA involvement in permethrin-induced cells is not clear enough.The purpose of this study was to investigate the regulatory role of miR-96 in the cell migration induced by low-dose permethrin, and to provide a new complement to the toxicity mechanism of permethrin. [Methods] By transfecting miR-96 analogs and inhibitors into human mammary gland cells MCF-7, the relative expression level of miR-96 was detected by fluorescence quantitative PCR, and miR-96 was artificially overexpressed and inhibited in the cells. The effect of permethrin on cell migration after miR-96 overexpression and inhibition was detected by cell scratch healing assay. [Results] The above experiments showed that the expression level of miR-96 in cells was positively correlated with the dose of permethrin. When treated with permethrin at 100 μmol/L for 48 h, miR-96 expression level was the highest(3.68±0.43), and the difference was extremely significant(P<0.01). After transfection of miR-96 analogues and inhibitors into MCF-7 cells, miR-96 expression was up-regulated by 2 fold and down-regulated by 4 fold, respectively.After 100 μmol/L permethrin was continued to be applied to cells, overexpression of miR-96 could enhance permethrin’s ability to promote cell migration and healing. Conversely, inhibition of miR-96 reduced this ability of permethrin. [Conclusions] Our study confirmed that miR-96 is involved in the migration of MCF-7 cells induced by permethrin, and permethrin is regulated by miRNA in the process of influencing cell physiological activities.