Effect Of Trastuzumab Combined With Nedaplatin On Proliferation Inhibition Of Ovarian Cancer SKOV3 Cells

Objective:To explore the effects of trastuzumab(Herceptin)combined with Nedaplatin on the proliferation of ovarian cancer SKOV3 cells and on the expression levels of P-AKT,AKT,Bcl-2 and Bax proteins.To provide the experimental and theoretical basis for the treatment of ovarian cancer.Method:Tetramethylazozo blue colorimetric method(MTT)was used to detect the proliferation inhibition of ovarian cancer SKOV3 cells after drug treatment;flow cytometry was used to detect the percentage change of the cell cycle of ovarian cancer SKOV3 cells after drug treatment;western Blotting was used to detect the effect of ovarian cancer SKOV3 cells on the protein expression levels of AKT,P-AKT,Bcl-2 and Bax after drug treatment.Result:1.MTT test results showed:Compared with the control group,the trastuzumab group(10μg/ml),nedaplatin group(1μg/ml、5μg/ml、10μg/ml、15μg/ml)and the combination group(1μg/ml、5μg/ml、10μg/ml、15μg/ml)could inhibit the proliferation of ovarian cancer SKOV3 cells(P<0.01);compared with the trastuzumab group,the inhibitory effect of the combination group was stronger(P<0.01);and the inhibitory effect of the combination group was stronger than the same concentration of nedaplatin group’s(P<0.05).2.The flow cytometry detection cycle results showed:(1)Compared with the control group,the trastuzumab group has no significant effect on the cell cycle distribution(P>0.05),(2)The effect on the G0/G1 phase distribution:Compared with the control group,the cell cycle distribution of G0/G1 phase in the nedaplatin group and the combination group were decreased(P<0.01);compared with the trastuzumab group,the cell cycle distribution of G0/G1 phase in the combination group was decreased(P<0.01);compared with the nedaplatin group,the cell cycle distribution of G0/G1 phase in the combination group was reduced(P<0.01);(3)The effect on the S phase distribution:Compared with the control group,the cell cycle distribution of S phase was decreased in the high concentration of nedaplatin combined group(P<0.05);compared with trastuzumab group,the cell cycle distribution of S phase in the high concentration of nedaplatin was decreased(P<0.01);compared with the high concentration of nedaplatin group,the cell cycle distribution of S phase in the high concentration of nedaplatin group was decreased(P<0.01);(4)The effect on the G2/M phase distribution:Compared with the control group,the low concentration of nedaplatin group had no significant effect on the G2/M phase(P>0.05);compared with the control group,the cell cycle distribution of G2/M phase in the high concentration of nedaplatin group,the low concentration of nedaplatin combination group and the high concentration of nedaplatin combination group were increased(P<0.01);compared with trastuzumab group,the cell cycle distribution of G2/M phase in the combination group was increased(P<0.01);compared with the nedaplatin group,the cell cycle distribution of G2/M phase in the combination group was increased(P<0.01).3.The results of Western blot analysis as follows:(1)The effect on the expression of AKT protein:Compared with the control group,The effect on the expression of AKT protein in trastuzumab group,nedaplatin group and combination were nonsignificant(P>0.05);(2)The effect on the expression of P-AKT protein:Compared with the control group,trastuzumab group had no significant effect on the expression of P-AKT protein(P>0.05);compared with the control group,the expression of P-AKT protein in low concentration of nedaplatin group and combined group were decreased(P<0.05);compared with trastuzumab group,the expression of P-AKT protein in combined group was decreased(P<0.05);compared with nedaplatin group,the expression of the P-AKT protein in the combined group was decreased(P<0.05);(3)The effect on the expression of Bcl-2 protein:Compared with the control group,the effect on the expression of Bcl-2 protein in the trastuzumab group and the low concentration of nedaplatin group were nonsignificant(P>0.05);compared with the control group,the expression of Bcl-2 protein in the high concentration of nedaplatin group and the combined group was reduced(P<0.05);compared with the trastuzumab group,the expression of Bcl-2 protein in the combination group were reduced(P<0.01);compared with the nedaplatin group,the expression of Bcl-2 protein in the the combination group were reduced(P<0.05);(4)The effect on the expression of Bax protein:Compared with the control group,the expression of Bax protein in the trastuzumab group and the low concentration of nedaplatin group were nonsignificant(P>0.05)compared with the control group,the expression of Bax protein in the high concentration of nedaplatin group and the combined group were increased(P<0.05);compared with trastuzumab group,the expression of Bax protein in the combination group were increased(P<0.01);compared with nedaplatin monotherapy group,the expression of Bax protein in the combination group were increased(P<0.05).Conclusion:1.Trastuzumab and nedaplatin have a synergistic effect on the proliferation inhibition of HER-2 positive ovarian cancer SKOV3 cells.2.Trastuzumab combined with nedaplatin has no significant effect on the expression of AKT protein,which can down-regulate the expression of P-AKT protein and Bcl-2 protein,and up-regulate the expression of Bax protein,thereby inducing apoptosis.3.Trastuzumab combined with nedaplatin prevents cell mitosis by blocking the cell cycle at the G2/M phase,thereby exerting an inhibitory effect on the proliferation of ovarian cancer SKOV3 cells.