GOLPH3 regulates proliferation and apoptosis of endometrial carcinoma cells through PI3K/AKT/GSK3β signal
10.3760/cma.j.issn.1673-422X.2020.02.001
- VernacularTitle:GOLPH3通过PI3K/AKT/GSK3β信号调控子宫内膜癌细胞的增殖与凋亡
- Author:
Chuncui LUO
1
;
Chaoyan YUAN
;
Qingfen CHEN
Author Information
1. 湖北民族大学附属民大医院妇科，恩施 445000
- From:
Journal of International Oncology
2020;47(2):65-69
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the mechanism of Golgi phosphoprotein 3 (GOLPH3) regulating the proliferation and apoptosis of endometrial cancer (EC) cells through PI3K/AKT/GSK3β signal.Methods:Human endometrial adenocarcinoma HEC-1-B cells were divided into control group, GOLPH3 group and GOLPH3 + GDC-0941 group. GOLPH3 was over-expressed by transfection of the GOLPH3 pcDNA. The PI3K inhibitor GDC-0941 was used to block the PI3K/AKT/GSK3β pathway. Cell viability, cell proliferation and apoptosis were measured by CCK-8 method, clone formation experiment and flow cytometry, respectively. The protein phosphorylation level in PI3K/AKT/GSK3β pathway was detected by Western blotting.Results:Transfection experiments were successful, and the PI3K inhibitor GDC-0941 did not affect GOLPH3 mRNA and protein expression. The relative cell viability of the control group, the GOLPH3 group and the GOLPH3 + GDC-0941 group were (100.00±4.63)%, (131.56±7.85)% and (97.85±7.36)%, and the difference among the three groups were significant ( F=7.437, P<0.001). The cell viability of the GOLPH3 group was higher than that of the control group ( P<0.001). The cell viability of the GOLPH3 + GDC-0941 group was lower than that of the GOLPH3 group ( P<0.001). The numbers of cell clones in the three groups were 46.86±3.56, 89.32±4.78 and 46.48±4.05, and the difference was significant ( F=20.437, P<0.001). GOLPH3 group had more clones than control group ( P<0.001). The number of clones formed in the GOLPH3 + GDC-0941 group was less than that in the GOLPH3 group ( P<0.001). The apoptosis rates of the three groups were (5.17±0.61)%, (2.34±0.56)% and (6.85±0.53)%, and the difference was significant ( F=11.643, P<0.001). The apoptosis rate of the GOLPH3 group was lower than that of the control group ( P<0.001), and the apoptosis rate of the GOLPH3 + GDC-0941 group was higher than that of the GOLPH3 group ( P<0.001). The phosphorylated PI3K/PI3K levels of the three groups were 1.00±0.09, 3.32±0.19 and 0.93±0.06, respectively; phosphorylated AKT/AKT levels were 1.00±0.11, 4.63±0.63 and 1.15±0.16, respectively; phosphorylated GSK3β/GSK3β levels were 1.00±0.08, 4.06±0.57 and 1.04±0.14, respectively. The differences were statistically significant ( F=12.532, P<0.001; F=16.792, P<0.001; F=15.311, P<0.001). The phosphorylation levels of each protein in the GOLPH3 group were higher than those in the control group (all P<0.001), and the GOLPH3 + GDC-0941 group were lower than the GOLPH3 group (all P<0.001). Conclusion:In EC cells, over-expression of GOLPH3 can promote cell proliferation and inhibit apoptosis by activating the PI3K/AKT/GSK3β pathway, suggesting that GOLPH3 is involved in the occurrence and development of EC.
