Abstract：Objective To investigate the effect and mechanism of miR-16 on proliferation and apoptosis of human breast cancer cells MDA-MB-231. Methods The cells cultured in vitro were divided into the following groups: human mammary epithelial cells HBL-100 (HBL-100 group ) and human breast cancer cells MDA-MB-231 (MDA-MB-231 group ). Real-time fluorescent quantitative PCR was used to detect miR-16 expression, and Western blot was used to detect β2 adrenergic receptor (β2-AR) protein expression. Transient transfection of cells was grouped as follows: MDA-MB-231 group, MDA-MB-231+miR-16 mimics (miR-16 mimics group), MDA-MB-231+miR-16 inhibitor (miR-16 inhibitor group), MDA-MB-231+ negative control (negative control group). The effects of miR-16 on proliferation and apoptosis of MDA-MB-231 cells were observed by MTT colorimetry and Annexin V-FITC double dye flow cytometry. The targeting relationship between miR-16 and β2-AR was verified by dual luciferase reporter genes. Results The expression of miR-16 in MDA-MB-231 group was lower than that in HBL-100 group, and the expression of β2-AR protein was higher than that in HBL-100 group, and the differences were statistically significant (P＜0.05). There was no significant difference in cell proliferation between MDA-MB-231 group and negative control group (P＞0.05). Cell proliferation in miR-16 mimics group was lower than that in MDA-MB-231 group, and cell proliferation in miR-16 inhibitor group was higher than that in MDA-MB-231 group, and the differences were statistically significant (P＜0.05). There was no significant difference in apoptosis rate between MDA-MB-231 group and negative control group (P＞0.05). The apoptosis rate in miR-16 mimics group was higher than that in MDA-MB-231 group, and the apoptosis rate in miR-16 inhibitor group was lower than that in MDA-MB-231 group, and the differences were highly statistically significant (P＜0.01). The results of dual luciferase reporter gene showed that, compared with MDA-MB-231 group, luciferase activity in miR-16 inhibitor group and negative control group had no significant changes, with no statistical significance (P＞0.05). Compared with MDA-MB-231 group, the luciferase activity of miR-16 mimics group with β2-AR 3’UTR sequence decreased, the difference was highly statistically significant (P＜0.01). Compared with MDA-MB-231 group, there was no significant change in luciferase activity in miR-16 mimics group with mutated β2-AR 3’UTR sequence, with no statistical significance (P＞0.05). Conclusion miR-16 may regulate the proliferation and apoptosis of breast cancer cells by targeting β2-AR.
刘嘉祺 周福波 林峰 付惠. miR-16对人乳腺癌细胞MDA-MB-231增殖和凋亡的作用及机制[J]. 中国医药导报, 2023, 20(25): 38-42.
LIU Jiaqi ZHOU Fubo LIN Feng FU Hui. Effect and mechanism of miR-16 on proliferation and apoptosis of human breast cancer cells MDA-MB-231. 中国医药导报, 2023, 20(25): 38-42.
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