miR-483靶向调控DLC1对三阴性乳腺癌细胞增殖、迁移和侵袭的影响

Effects of miR-483 targeting DLC1 on the proliferation,migration and invasion of triple negative breast cancer cells

目的探讨miR-483靶向调控肝癌缺失基因1(DLC1)对三阴性乳腺癌细胞增殖、迁移和侵袭的影响。方法将MDA-MB-231乳腺癌细胞分为对照组、miR-NC组、miR-483 inhibitor组、miR-483 mimics组,各组细胞设6个平行样,培养72 h。采用MTT法检测各组细胞活力,结晶紫染色检测细胞单克隆形成数目,流式细胞仪测定细胞凋亡水平,Transwell小室测定细胞迁移与侵袭水平,RT-qPCR法及蛋白印迹法测定细胞miR-483、DLC1 mRNA和蛋白表达水平。结果与对照组、miR-NC组比较,miR-483 inhibitor组细胞OD值、存活率、单克隆形成数目、穿膜数、miR-483 mRNA表达水平降低/减少(P<0.05),miR-483 mimics组细胞OD值、存活率、单克隆形成数目、穿膜数、miR-483 mRNA表达水平升高/增加(P<0.05);与miR-483 inhibitor组比较,miR-483 mimics组OD值、存活率、单克隆形成数目、穿膜数、miR-483 mRNA表达水平升高/增加(P<0.05)。与对照组、miR-NC组比较,miR-483 inhibitor组凋亡率、DLC1 mRNA和蛋白表达水平升高(P<0.05),miR-483 mimics组凋亡率、DLC1 mRNA和蛋白表达水平降低(P<0.05);与miR-483 inhibitor组比较,miR-483 mimics组凋亡率、DLC1 mRNA和蛋白表达水平降低(P<0.05)。结论miR-483过表达能促进三阴性乳腺癌细胞增殖、侵袭,抑制其凋亡,其机制可能与miR-483抑制DLC1表达有关。

Objective To investigate the effects of miR-483 targeting deleted in liver cancer 1(DLC1)on the proliferation,migration and invasion of triple negative breast cancer cells.Methods The MDA-MB-231 breast cancer cells were divided into the control group,the miR-NC group,the miR-483 inhibitor group,and the miR-483 mimics group.Six parallel cells were set up in each group and cultured for 72 hours.MTT assay was used to detect the cell viability of each group,crystal violet staining was used to detect the number of monoclonal formation of cells,flow cytometry was used to determine the level of cell apoptosis,Transwell chamber was used to determine the level of cell migration and invasion,and RT-qPCR and Western blot were used to determine the expression levels of miR-483,DLC1 mRNA and protein in cells.Results Compared with the control group and the miR-NC group,the cell absorbance(OD)value,survival rate,number of monoclonal formation,number of penetrating membrane,and the expression level of miR-483 mRNA in the miR-483 inhibitor group decreased(P<0.05);the OD value,survival rate,number of monoclonal formation,number of penetrating membrane,and the expression level of miR-483 mRNA in the miR-483 mimics group increased(P<0.05).Compared with the miR-483 inhibitor group,the OD value,survival rate,number of monoclonal formation,number of penetrating membrane,and the expression level of miR-483 mRNA in the miR-483 mimics group increased(P<0.05).Compared with the control group and the miR-NC group,the apoptosis rate,the expression levels of DLC1 mRNA and protein in the miR-483 inhibitor group increased(P<0.05),and the apoptosis rate,the expression levels of DLC1 mRNA and protein in the miR-483 mimics group decreased(P<0.05);compared with the miR-483 inhibitor group,the apoptosis rate,the expression levels of DLC1 mRNA and protein in the miR-483 mimics group decreased(P<0.05).Conclusion The overexpression of miR-483 can promote the proliferation and invasion of triple negative breast cancer cells and inhibit their apoptosis;the mechanism may be related to the inhibition of DLC1 expression by miR-483.