The Mechanism Of Apoptosis Of Human Ovarian Cancer SKOV3 Cells Induced By PTEN Via PI3K/AKT/NF-κB Signaling Pathway

Objective The purpose of this study was to investigate the role of PTEN and its related pathways in human ovarian serous carcinoma SKOV3 cells and its related mechanisms.Methods Human ovarian serous carcinoma SKOV3 cell line with PTEN gene overexpression was constructed by liposome transfection method,and verified by Real-time PCR and Western blot.The in vitro proliferation of SKOV3 cells before and after PTEN gene overexpression was detected by CCK 8 assay and clonal formation assay.The effects of PTEN gene overexpression on the migration ability of SKOV3 cells in vitro were detected by cell scratch and Transwell assay.Hoechst/PI double staining technique and flow cytometry were used to detect the apoptosis of SKOV3 cells before and after PTEN gene overexpression.The expression levels of PI3K/AKT/NF-кB pathway proteins and apoptosis proteins in SKOV3 cells before and after PTEN gene overexpression by Western blot.Results PTEN m RNA and protein levels of PTEN gene overexpression group were significantly higher than those of blank control group and negative control group（RT-PCR:F=16.352,P=0.004;Western blot:F=24.665,P=0.001）.The results of CCK8 and clonal formation assay showed that the cell proliferation level of PTEN overexpression group was significantly decreased in the PTEN overexpression group compared with the blank control group and the negative control group（CCK8 assay:F=210.885,P=0.000;Clone formation experiment:F=52.659,P=0.000）.The results of cell scratch and Transwell assay showed that the level of cell migration in the PTEN overexpression group was significantly decreased compared with the blank control group and the negative control group（cell scratch method:F=60.358,P=0.000;Transwell method:F=16.803,P=0.000）.Hoechst/PI staining showed that the apoptosis rate of PTEN overexpression group was significantly increased compared with blank control group and negative control group（F=10.364,P=0.011）.Flow cytometry showed that the apoptosis rate of PTEN overexpression group was significantly higher than that of blank control group and negative control group（F=23.916,P=0.001）.Western blot results showed that the expression of p-PI3K/PI3K,p-AKT/AKT,NF-кB,IKKβand BCL-2/Bax proteins was significantly reduced in the PTEN gene overexpression group compared with the blank control and negative control groups,and the differences were statistically significant(F_p-PI3K/PI3K=15.982,P=0.004;F_p-AKT/AKT=29.788,P=0.001;F_NF-кB=19.741,P=0.002;F_IKKβ=11.799,P=0.008;F_BCL-2/Bax=17.719;P=0.003).Conclusion The proliferation level and migration ability of SKOV3 cells in ovarian serous carcinoma with overexpression of PTEN gene are inhibited,and the apoptosis rate of cells increased,suggesting that PTEN may promote apoptosis of tumor cells by inhibiting PI3K/AKT/NF-кB signaling pathway,thus inhibiting the proliferation and migration ability of tumor cells.