Effect and mechanism of miR-195 regulates on proliferation and apoptosis of human breast cancer MCF-7 cells#br#
LIU Jiaqi1 ZHAI Fengguo1 GAO Jinxia2 LIU Jiawei3 YANG Xudong1 ZHOU Fubo1 LI Mengquan4▲
1.Department of Pharmacology, Mudanjiang Medical University, Heilongjiang Province, Mudanjiang 157011, China;
2.Department of Preventive Medicine, Mudanjiang Medical University, Heilongjiang Province, Mudanjiang 157011, China;
3.Department of Organic Chemistry, Mudanjiang Medical University, Heilongjiang Province, Mudanjiang 157011, China;
4.Medical Research Center, Mudanjiang Medical University, Heilongjiang Province, Mudanjiang 157011, China
Abstract:Objective To explore the role and mechanism of miR-195 in regulating proliferation and apoptosis of human breast cancer MCF-7 cells (MCF-7). Methods Breast epithelial cells MCF-10A (MCF-10A) and MCF-7 were cultured in vitro. The expression of miR-195 was detected by qRT-PCR. Protein expression of phosphatase magnesium-dependent 1 delta (PPM1D) was detected by Western blot (WB). Transient transfection of MCF-7 was carried out. 3-(4,5- two methyl thiazole -2)-2,5- two phenyl tetrazolium bromide (MTT) method and DNA breakage TdT-mediated-duTP nick end labeling method were used to observe the effect of miR-195 on proliferation and apoptosis of MCF-7. Double luciferase reporter genes were used to verify the relationship between miR-195 and PPM1D. Results The expression of miR-195 in MCF-7 group was lower than that in MCF-10A group, the expression of PPM1D protein in MCF-7 group was higher than that in MCF-10A group, the differences were statistically significant (P < 0.05). The cell viability of miR-195 mimics group was lower than MCF-7 group; miR-195 inhabitor group was higher than MCF-7 group, the differences were highly statistically significant (P < 0.01). The apoptosis rate of miR-195 mimics group was higher than MCF-7 group, the difference was highly statistically significant (P < 0.01); and miR-195 inhabitor group was lower than MCF-7 group, the difference was statistically significant (P < 0.05). Double luciferase reporter gene showed that miR-195 mimics group, which carried the PPM1D gene fragment, the activity of luciferase was lower than MCF-7 group , the difference was statistically significant (P < 0.01). There was no significant difference in luciferase activity between the mir-195 MIMics group containing the mutated PPM1D gene fragment and McF-7 group (P > 0.05). Conclusion miR-195 may regulate the proliferation and apoptosis of breast cancer cells through targeting PPM1D.
刘嘉祺1 翟凤国1 高金霞2 刘佳维3 杨旭东1 周福波1 李孟全4▲. miR-195调控人乳腺癌细胞MCF-7增殖与凋亡的作用及机制研究[J]. 中国医药导报, 2022, 19(1): 18-22.
LIU Jiaqi1 ZHAI Fengguo1 GAO Jinxia2 LIU Jiawei3 YANG Xudong1 ZHOU Fubo1 LI Mengquan4▲. Effect and mechanism of miR-195 regulates on proliferation and apoptosis of human breast cancer MCF-7 cells#br#. 中国医药导报, 2022, 19(1): 18-22.
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