MIR-129 INHIBITS GLIOMA CELL PROLIFERATION AND PROMOTES AUTOPHAGY AND APOPTOSIS BY TARGETING NOTCH-1

Gang Zong, Xiuhao Huo, Tian Huai, Erbao Bian, Bing Zhao^* 

Department of Neurosurgery, the Second Affiliated Hospital of Anhui Medical University, Hefei, PR China

Department of Neurosurgery, the Second Affiliated Hospital of Anhui Medical University, Hefei, PR China

Objective: To analyze the inhibitory effect of miR-129 on glioma cell proliferation, autophagy, and apoptosis by targeting Notch-1. 

Methods: Human glioma cell line U251 was cultured and subcultured, then divided into a control group, a miR-129 overexpression group, and a miR-129 inhibition group. U251 cells were transfected with a pE2F7 overexpression vector and an interference vector, and the blank control group was treated with a blank lentivirus vector. MTT was used to detect the cell growth curve; flow cytometry was used to detect the apoptosis rate; real-time PCR was used to detect the expression of LC3-Ⅱ/LC3-Ⅰ, p62, and Notch-1 mRNA; and Western blot was used to detect the protein expression of LC3-Ⅱ/LC3-Ⅰ, p62, and Notch-1. 

Results: The proliferation of the three groups increased with time. The proliferation of the miR-129 overexpression group was significantly lower than that of the control group at day 3 through day 7, and the cell proliferation in the miR-129 inhibition group was significantly higher than that of the control group (P<0.05). The apoptosis rate of the miR-129 group was significantly higher than that of the control group, and the apoptosis rate of the miR-129 inhibition group was significantly lower than that of the control group (P<0.05). The expression levels of LC3 - Ⅱ/LC3 - Ⅰ protein and mRNA in the miR-129 group were significantly higher than those in the control group; p62 protein and mRNA expression levels were significantly lower in the miR-129 group than they were in the control group; and p62 protein and mRNA expression levels in the miR-129 group were significantly lower than they were in the control group (P<0.05). The expression levels of Notch-1 protein and mRNA in the miR-129 group were significantly lower than those in the control group, and the expression levels of Notch-1 protein and mRNA in the miR-129 group were significantly higher than those in the control group (P<0.05). 

Conclusion: miR-129 can inhibit the proliferation of glioma cells, increase the apoptosis rate of glioma cells, and promote autophagy by regulating the expression of autophagy-related proteins. The mechanism may be related to miR-129 regulating Notch-1.