Role of ATR/CHK1 pathway in the occurrence and development of epithelial ovarian cancer / 中华内分泌外科杂志

ABSTRACT

Objective:To discuss the role of ataxia telangiectasia and Rad3-related kinase (ATR) /check point kinase 1 (CHK1) signaling pathway in the occurrence and development of epithelial ovarian cancer.Methods:Human epithelial ovarian cancer cells OVCAR3 cells were cultured in vitro, siRNA and VE-822 were used to interfere with ATR in OVCAR3 cells, the effectiveness of interference with ATR expression was detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, the cell proliferation inhibition was detect by CCK-8 method, cell cycle and apoptosis were detected by flow cytometry (FCM) , the mRNA expressions of Caspase-3, B-cell lymphoma-2 (Bcl-2) , Bcl-2-associated X protein (Bax) were detected by qRT-PCR, Western blot was used to detect the expression of ATR/CHK1 pathway related proteins.Results:After siRNA-ATR transfection and VE-822 intervention, compared with those in NC group and siRNA-sc group, the expression level of ATR mRNA [ (1.55±0.12) , (1.51±0.13) , (0.71±0.11) , (0.73±0.12) , (0.49±0.09) ] in OVCAR3 cells in siRNA-ATR group, VE-822 group and siRNA-ATR + VE-822 group was significantly lower ( P<0.05) , the inhibition rate of cell proliferation [ (0.00±0.00) %, (0.00±0.00) %, (32.84±1.08) %, (30.75±1.44) %, (43.90±1.57) %], ratio of G0/G1 phase [ (40.08±2.57) , (36.35±3.44) , (53.28±4.34) , (56.37±5.03) , (70.63±3.81) ], apoptosis rate [ (4.28±0.67) %, (5.35±0.94) %, (23.63±1.13) %, (24.57±1.20) %, (35.86±1.09) %], expression of Caspase-3 and Bax mRNA were significantly higher ( P<0.05) , and the cell proportions in S phase [ (32.93±3.02) , (35.35±2.82) , (25.79±3.61) , (23.74±3.54) , (18.04±2.37) ] and G2/M phase [ (26.99±2.84) , (28.30±2.72) , (20.93±3.01) , (19.98±2.87) , (11.33±2.11) ], Bcl-2 mRNA, ATR, p-CHK1/CHK1, cell division cycle protein 25C (CDC25C) and cyclin B1 protein expression were significantly lower ( P<0.05) . Compared with those in siRNA-ATR group, the expression of ATR mRNA in siRNA-ATR + VE-822 group was further decreased ( P<0.05) , the inhibition rate of cell proliferation, apoptosis, expression of Caspase-3 and Bax mRNA were further increased ( P<0.05) , and the expression of Bcl-2 mRNA, ATR, p-CHK1/CHK1, CDC25C and cyclin B1 protein was decreased continuously ( P<0.05) . Conclusion:ATR/CHK1 signaling pathway is activated during the proliferation of epithelial ovarian cancer OVCAR3 cells. Inhibition of ATR/CHK1 signaling pathway can inhibit the proliferation of epithelial ovarian cancer cells and induce G1/S cell cycle arrest and apoptosis.