miR-944过表达对三阴乳腺癌细胞系MDA-MB-231的侵袭迁移和上皮间质转化抑制作用及其机制

Inhibitory effects of miR-944 overexpression on invasion,migration,and epithelial mesenchymal transi⁃tion in triple-negative breast cancer cell line MDA-MB-231

目的探讨miR-944过表达对三阴乳腺癌(TNBC)MDA-MB-231细胞系的侵袭、迁移和上皮间质转化(EMT)的抑制作用及其可能的作用机制。方法采用qRT-PCR法检测TNBC细胞系MDA-MB-231、MDA-MB-453、HCC1806和正常乳腺上皮细胞系HBL-100中的miR-944。选取miR-944相对表达量最低的MDA-MB-231进行后续实验。收集对数生长期的MDA-MB-231分为miR-944过表达组(miR-944-IN组)、阴性对照组(miR-944-NC组),miR-944-IN组细胞转染miR-944 mimics,miR-944-NC组细胞转染NC-mimics(模拟物对照)。转染成功后,应用细胞划痕实验检测两组细胞迁移能力,Transwell侵袭实验检测细胞侵袭能力,Westernblotting法检测EMT相关标志物E-cadherin、Fibronectin、Vimentin蛋白及GATA6蛋白。结果与miR-944-NC组相比,miR-944-IN组MDA-MB-231细胞中miR-944的相对表达量升高(P<0.05),细胞划痕愈合率降低(P<0.05),穿膜细胞数减少(P<0.05),E-cadherin蛋白相对表达量升高(P<0.05),Fibronectin、Vimentin、GATA6蛋白相对表达量降低(P均<0.05)。结论miR-944在TNBC细胞中表达降低。过表达miR-944能够抑制TNBC细胞的迁移、侵袭及EMT,其机制可能与抑制GATA6蛋白有关。

Objective To investigate the inhibitory effects of overexpression of miR-944 on invasion,migration,and epithelial-mesenchymal transition(EMT)of triple-negative breast cancer(TNBC)and the possible mechanism.Methods The qRT-PCR was used to detect miR-944 in TNBC cell lines MDA-MB-231,MDA-MB-453,HCC1806,and normal breast epithelial cell line HBL-100.MDA-MB-231 with the lowest expression level of miR-944 was selected for fol-low-up experiments.MDA-MB-231 cells in the logarithmic phase were collected and divided into the miR-944 overexpres-sion group(miR-944-IN group)and negative control group(miR-944-NC group).Cells in the miR-944-IN group were transfected with miR-944 mimics and cells in the miR-944-NC group were transfected with NC-mimics,respectively.After the successful transfection,the cell migration ability of the two groups was detected by Scratch assay,the cell invasion abil-ity was detected by Transwell invasion test,and the EMT-related markers E-cadherin,Fibronectin,Vimentin protein and GATA6 protein were detected by Western blotting.Results Compared with the miR-944-NC group,the relative expres-sion of miR-944 in MDA-MB-231 cells increased(P<0.05),the wound healing rate decreased(P<0.05),the number of transmembrane cells decreased significantly(P<0.05),the expression level of E-cadherin protein increased(P<0.05),and the expression levels of Fibronectin,Vimentin and GATA6 protein decreased in the miR-944-IN group(all P<0.05).Conclusions The expression of miR-944 decreases in TNBC cells.Overexpression of miR-944 can inhibit the migra-tion,invasion and EMT of TNBC cells,and its mechanism may be related to its inhibition of GATA6 gene.