Mechanism Of Combined Inhibitory Effect Of Trastuzumab With Lobaplatin On Ovarian Cancer SKOV-3 Cells

Objectives:To explore the inhibitory effects of the Trastuzumab(Herceptin)combined with the Lobaplatin(lob)on the HER-2-positive ovarian cancer SKOV-3 cell lines and explore the underlying mechanism.Methods:Ovarian cancer cells SKOV-3 was cultured in the vitro with the McCoy’s 5 A culture medium and the morphology and growth pattern of the cells were observed under the electron microscope.The control group,trastuzumab(10μg/ml)group,lobaplatin(1μg/ml)group,and the combined group(trastuzumab10μg/ml+lobaplatin1μg/ml)were treated with dose and observed.Methyl thiazolylterazolium(MTT)was used to determine the effects of drug on cell proliferation inhibition,and the inhibitory rate of drug.Cell cycle distribution of each group has been determined by the flow cytometry tests.The western blotting was used for the protein expression of AKT,Bcl-2,P53,and Bax in SKOV-3 cells.Results:1.The MTT results showed that the Trastuzumab with the concentration of 0.1μg/ml,1μg/ml,10μg/ml,and 100μg/ml had no cytotoxic effects on ovarian cancer SKOV-3 cells for 24hrs,48hrs,72hrs,and 96hrs respectively;Compared with the control group,trastuzumab group(10μg/ml)was non-toxic(P>0.05),while lobaplatin group(1μg/ml)was slightly toxic to cell(P>0.05).The CI of the two drugs is 1.2,which belongs to synergism.2.The cell cycle distribution by flow cytometry result shows that:(1)Compared wit h the control group,Trastuzumab alone had no significant effect on cell cycle distribu tion of SKOV-3 cells(P>0.05),and the distribution proportion of G0/G1 phase in SKOV-3 cells was slightly decreased(P>0.05),while trastuzumab combined with Lobaplatin significantly reduced the G0 of SKOV-3 cells.There was statistical significance in the distribution G1 phase in SKOV-3 cells(P<0.05).(2)Compared with the control group,the trastuzumab group of cell cycle distribution had no significant effect on the SKOV-3 cells(P>0.05),and the proportion of S-phase distribution in SKOV-3 cells was slightly reduced in lobaplatin group(P>0.05),while trastuzumab combined with the lobaplatin group has reduced the distribution ratio on S-phase in SKOV-3 cells(P<0.05).(3)Compared with the Control group,trastuzumab has no significant effect on G2/M phase distribution ratio of SKOV-3 cells(P>0.05),but the G2/M phase distribution ratio of SKOV-3 cells in the lobaplatin group was slightly Reduced(P>0.05),and the SKOV-3 cell distribution ratio in the combined treatment group was significantly increased(P<0.05).These results suggested that the trastuzumab combined with Lobaplatin can cause the arrest of cell cycle distribution.3.The results of Western blotting analysis showed that:(1)Protien expression of p-AKT;Compared with the control group,the protein expression of p-AKT was slightly decreased in the trastuzumab group(P>0.05),and in the lobaplatin group(P>0.05),the expression of p-AKT was significantly decreased in the combined treatment group(P<0.05)(2)The protein expression of wt-p53:Compared with the control group,the protein expression of the trastuzumab group was slightly decreased(P>0.05),while theprotein expression of lobaplatin group was slightly reduced(P>0.05),and the combined treatment groupwas slightly enhanced(P<0.05)(3)the protein expression of Bax:compared with the control group,the expression trastuzumab group had no significant effect(P>0.05),while the lobaplatin group was slightly enhanced(P>0.05),and the expression of the combined treatment group was significantly enhanced(P<0.05).(4)Bcl-2 expression:compared with the control group,the expression the trastuzumab group had no significant effect(P>0.05),while the protein expression of the lobaplatin group was slightly reduced(P>0.05),the expression of combined treatment group was decreased,and the difference was statistically significant(P<0.05).Conclusion:1.Trastuzumab combined with lobaplatin may cause cytotoxic effects on human ovarian cancer SKOV-3 cells by inhibiting the cell cycle.2.Trastuzumab and lobaplatin may inhibit the proliferation of tumor by down-regulation of P-AKT,Bcl-2 and upregulating the expression of Bax,and p53,and induce apoptosis of the tumor cell.3.The combination of Trastuzumab and lobaplatin maybe a new clinical treatment model for ovarian cancer.