Proliferation and invasion of breast cancer cells by targeting microRNA-1246 to StarD13
Objective To investigate the molecular mechanism of miR-1246 promoting the proliferation,migration and inva-sion of breast cancer cells.Methods The expression of miR-1246 and its relationship with the survival rate of breast cancer patients were analyzed by bioinformatics.Real-time fluorescence quantitative polymerase chain reaction(qRT-RCR)was used to evaluate the expression of miR-1246 in breast cancer cells and breast cancer tissues.Transwell chamber was used to detect cell invasion ability;EdU assay and cloning assay were used to detect cell proliferation.Scratch test was used to detect cell migration ability;the expression and survival curve of StarD13,the target protein of miR-1246,in breast cancer were determined by bioinformatics.The relationship between miR-1246 and StarD13 was detected by dual luciferase reporter gene.Western blot was used to detect the expression of related target protein StarD13 after down-regu-lation of miR-1246.Independent sample t test was used to analyze the quantitative data between the two groups,and one-way analysis of variance was used to compare multiple groups.Results miR-1246 was highly expressed in breast cancer tissues and breast cancer cell MDA-MB-231.Transwell assay showed that the invasion ability of miR-1246 inhibitor group was lower than that of NC inhibitor group(t=9.14,P＜0.01).The results of EdU experiment showed that the cell proliferation ability decreased after down-regulation of miR-1246(t=6.81,P＜0.01).Colony formation assay showed that the proliferation ability of cells decreased after down-regulation of miR-1246(t=5.41,P＜0.01).The results of scratch test showed that the cell migration rates of miR-1246 inhibitor and NC inhibitor groups were 0.21±0.02 and 0.50 ±0.04,respectively,and the difference was statistically significant(t=13.13,P＜0.01).Bioinformatics predicted that there was a complementary sequence between miR-1246 and StarD13;breast cancer patients with low expression of StarD13 in breast cancer tissues and low expression of StarD13 had a poor prognosis(P＜0.01).The expression of StarD13 was increased when miR-1246 inhibitor was transfected.Dual-luciferase reporter assay confirmed that StarD13 could be used as a target gene of miR-1246.At the same time,the results of EdU assay showed that the positive rate of EdU in miR-1246 inhibitor+siStarD13 group[(55.89±1.95)％]was higher than that in miR-1246 inhibitor+NC group[(25.24±6.17)％],and the difference was statistically significant(t=8.21,P＜0.01).Trans well assay showed that compared with the miR-1246 inhibitor+NC group(127.67±23.46),the number of cells passing through the basement membrane in the miR-1246 inhibitor+siStarD13 group(385.67±11.59)was significantly increased,and the difference was statistically significant(t=17.08,P＜0.001).Conclusion miR-1246 can promote the proliferation,migration and invasion of breast cancer cells by targeting StarD13.
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