GOLPH3 regulates proliferation and apoptosis of endometrial carcinoma cells through PI3K/AKT/GSK3β signal

Abstract

Abstract:

ObjectiveTo explore the mechanism of Golgi phosphoprotein 3 (GOLPH3) regulating the proliferation and apoptosis of endometrial cancer (EC) cells through PI3K/AKT/GSK3β signal.MethodsHuman 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.ResultsTransfection 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 (allP<0.001), and the GOLPH3 + GDC-0941 group were lower than the GOLPH3 group (allP<0.001).ConclusionIn 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.

Key words:Endometrial neoplasms,Cell proliferation,Apoptosis,Golgi phosphoprotein 3,PI3K/AKT

Luo Chuncui, Yuan Chaoyan, Chen Qingfen. GOLPH3 regulates proliferation and apoptosis of endometrial carcinoma cells through PI3K/AKT/GSK3β signal[J]. Journal of International Oncology, 2020, 47(2): 65-69.

Figures/Tables4

组别	GOLPH3 mRNA	GOLPH3蛋白
对照组	1.00±0.08	1.00±0.10
GOLPH3组	4.67±0.32^a	3.16±0.29^a
GOLPH3+GDC-0941组	4.78±0.37^a	3.14±0.40^a
F值	17.574	12.683
P值	<0.001	<0.001

组别	p-PI3K/PI3K	p-AKT/AKT	p-GSK3β/GSK3β
对照组	1.00±0.09	1.00±0.11	1.00±0.08
GOLPH3组	3.32±0.19^a	4.63±0.63^a	4.06±0.57^a
GOLPH3+GDC-0941组	0.93±0.06^b	1.15±0.16^b	1.04±0.14^b
F值	12.532	16.792	15.311
P值	<0.001	<0.001	<0.001

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