miR-101-3p靶向调控Rac1对乳腺癌细胞侵袭和迁移的影响

Effects of miR-101-3p on invasion and migration of breast cancer cells by targeting Rac1

目的探讨miR-101-3p靶向调控Rac1对乳腺癌细胞侵袭和迁移的影响。方法体外培养乳腺癌细胞MCF-7、SK-BR-3、MDA-MB-435及正常乳腺细胞HBL-100,采用qRT-PCR法检测各细胞miR-101-3p表达。选择miR-101-3p表达最低的乳腺癌细胞进行后续实验。将miR-101-3p表达最低的乳腺癌细胞随机分为对照组、miR-101-3p抑制物组和miR-101-3p mimics组,miR-101-3p抑制物组和miR-101-3p mimics组分别转染miR-101-3p抑制物、miR-101-3p mimics。转染24 h,收集细胞,采用Transwell侵袭和迁移实验检测细胞侵袭和迁移能力。将miR-101-3p表达最低的乳腺癌细胞随机分为野生型Rac1+miR-101-3p mimics组、突变型Rac1+miR-101-3p mimics组、野生型Rac1+阴性对照物组、突变型Rac1+阴性对照物组,分别加入相应的转染物和转染试剂。转染24 h,收集细胞,采用双荧光素酶报告基因实验检测各组荧光素酶活性。结果乳腺癌MCF-7、SK-BR-3、MDA-MB-435细胞miR-101-3p相对表达量均明显低于正常乳腺HBL-100细胞(P均<0.05),且以MDA-MB-435细胞miR-101-3p相对表达量最低。miR-101-3p抑制物组细胞侵袭和迁移能力均明显高于对照组,而miR-101-3p mimics组细胞侵袭和迁移能力均明显低于对照组(P均<0.05)。野生型Rac1+miR-101-3p mimics组荧光素酶活性明显低于其他三组(P均<0.05),其他三组两两比较P均>0.05。结论 miR-101-3p可能通过靶向调控Rac1抑制乳腺癌细胞的侵袭和迁移。

Objective To investigate the effects of miR-101-3p on the invasion and migration of breast cancer cells by targeting Rac1.Methods Breast cancer cells MCF-7,SK-BR-3,MDA-MB-435 and normal breast cells HBL-100 were cultured in vitro.The expression of miR-101-3p in the breast cancer cells was detected by qRT-PCR.The breast cancer cells with the lowest expression of microRNA-101-3p were selected for subsequent experiments.Breast cancer cells were randomly divided into the control group,miR-101-3p inhibitor group,and miR-101-3p mimics group.The cells in the MiR-101-3p inhibitor group and miR-101-3p mimics group were transfected with miR-101-3p inhibitor and miR-101-3p mimics,respectively.After 24-hour transfection,the cells were collected and their invasion and migration abilities were tested by Transwell invasion and migration experiments.Breast cancer cells were randomly divided into the wild-type Rac1+miR-101-3p mimics group,mutant Rac1+miR-101-3p mimics group,wild-type Rac1+negative control group,and mutant Rac1 wild-type+negative control group,and the corresponding transfectants and transfection reagents were added,respectively.After 24-hour transfection,the cells were collected and the luciferase activity of each group was detected by double luciferase reporter gene assay.Results The relative expression of miR-101-3p in the breast cancer MCF-7,SK-BR-3 and MDA-MB-435 cells was lower than that in the normal breast HBL-100 cells(all P<0.05),and the relative expression of miR-101-3p in the MDA-MB-435 cells was the lowest,so the subsequent experiments were carried out with MDA-MB-435 cells.The results of Transwell invasion and migration experiment showed that the invasion and migration abilities of the cells in the miR-101-3p inhibitor group were significantly higher than those in the control group,while the invasion and migration abilities of the cells in the miR-101-3p mimics group were significantly lower than those in the control group(all P<0.05).The luciferase activity of wild-type Rac1+miR-101-3p mimics group was significantly lower than that of the other three groups(all P<0.05),and no significant difference was found among the other three groups(all P>0.05).The results confirmed that Rac1 was the direct target gene of miR-101-3p.Conclusion MiR-101-3p may regulate the invasion and migration of breast cancer cells by targeting Rac1.