雷公藤内酯醇通过调控miR-142-3p/HSP70通路抑制人乳腺癌MCF-7细胞增殖、侵袭和迁移

Triptolide inhibits proliferation,invasion and migration of human breast cancer MCF-7 cells by regulating miR-142-3p/HSP70 pathway

目的:探究雷公藤内酯醇(TP)通过miR-142-3p/HSP70信号通路对人乳腺癌MCF-7细胞恶性生物学行为的影响。方法:常规培养MCF-7细胞,将其分为6组:对照组、TP组、miR-142-3p inhibitor组、TP+inhibitor组、miR-142-3p mimic组和TP+mimic组,用转染试剂将相应的核酸或质粒转染MCF-7细胞。qPCR法、EdU细胞增殖实验、Transwell小室实验、细胞划痕实验、WB法分别检测转染后各组MCF-7细胞中miR-142-3p和HSP70 mRNA的表达,MCF-7细胞的增殖、侵袭、迁移能力和HSP70蛋白表达水平。结果:TP或miR-142-3p过表达能显著促进MCF-7细胞中miR-142-3p和HSP70的表达,敲减miR-142-3p则可明显抑制MCF-7细胞中miR-142-3p和HSP70的表达,TP可逆转由敲减miR-142-3p对MCF-7细胞中miR-142-3p和HSP70表达的影响;TP、过表达miR-142-3p均可明显抑制MCF-7细胞的增殖、迁移和侵袭能力(均P<0.05),敲减miR-142-3p则均可促进MCF-7细胞的增殖、迁移和侵袭能力(均P<0.05),TP可逆转由敲减miR-142-3p对MCF-7细胞恶性生物学行为的影响(均P<0.05)。结论:TP可通过调控miR-142-3p/HSP70信号通路,进而抑制MCF-7细胞的增殖、侵袭和迁移能力。

Objective:To investigate the effects of triptolide(TP)on the malignant biological behaviors of human breast cancer MCF-7 cells through the miR-142-3p/HSP70 signaling pathway.Methods:MCF-7 cells were routinely cultured and divided into 6 groups:control group,TP group,miR-142-3p inhibitor group,TP+inhibitor group,miR-142-3p mimics group and TP+mimics group.The corresponding nucleic acids or plasmids were transfected into MCF-7 cells with transfection reagents.The mRNA expression of miR-142-3p and HSP70 in MCF-7 cells was detected by qPCR method.The proliferation,invasion and migration abilities of MCF-7 cells were detected by EdU cell proliferation assay,Transwell chamber assay,and cell scratch assay,respectively.The protein expression of HSP70 in MCF-7 cells was determined by WB assay.Results:TP or miR-142-3p overexpression significantly promoted the expression of miR-142-3p and HSP70 in MCF-7 cells,while knockdown of miR-142-3p significantly inhibited the expression of miR-142-3p and HSP70 in MCF-7 cells.TP could reverse the effects of miR-142-3p knockdown on the expression of miR-142-3p and HSP70 in MCF-7 cells.TP and miR-142-3p overexpression could significantly inhibit the proliferation,migration and invasion of MCF-7 cells(all P<0.05),while knockdown of miR-142-3p could promote the proliferation,migration and invasion of MCF-7 cells(all P<0.05).TP could reverse the effect of miR-142-3p knockdown on the malignant biological behavior of MCF-7 cells(all P<0.05).Conclusions:TP can inhibit the proliferation,invasion,and migration of MCF-7 cells by regulating the miR-142-3p/HSP70 signaling pathway.