| Objectives:To investigate the effects of trastuzumab combined with oxaliplatin on the activity of HER2-positive ovarian cancer cells and relevant signaling pathways,including p-AKT,p-ERK,Bcl-2 and Bax,as well as to explore the possibility and effectiveness of the combination of trastuzumab and oxaliplatin.Materials and methods:Human ovarian cancer cell line SKOV3 cells were cultured in vitro.Four groups were set:control group,oxaliplatin group,trastuzumab group,combination group(oxaliplatin and trastuzumab),and the foliowing experiments were carried out:1.Morphological observation of the drug-treated cells under optical micros copy;2.MTT assay was used to determine the effects of drugs on cell proliferative inhibition,and the inhibition rate of drugs on cell proliferation was analyzed;3.Flow cytometry was used to detect cell cycle;4.The protein expression of p-AKT,p-ERK,Bcl-2 and Bax in SKOV3 cells was detected by Western blot.Results:1.Different concentrations of trastuzumab(0.1μg/ml,1μg/ml,10μg/ml,100μg/ml)were not cytotoxic to SKOV3 cells after incubation for 24h,48h,72h and 96h,respectively(P>0.05).2.Cell cycle of SKOV3 by flow cytometry showed:(1)Compared with the control group,trastuzumab alone failed to have significant effects on the cell cycle distribution of SKOV3 cells(P>0.05),oxaliplatin alone decreased the distribution ratio of G0/G1 phase in SKOV3 cells(P<0.05).The combination of two drugs significantly reduced the distribution ratio of G0/G1 phase in SKOV3 cells(P<0.01).Compared with the trastuzumab group,the distribution ratio of G0/G1 phase was decreased in SKOV3 cells in the combination group(P<0.01).Compared with the oxaliplatin group,the distribution ratio of G0/G1 phase in SKOV3 cells was decreased in the combination group(P<0.05).(2)Compared with the control group,the S phase distribution ration of SKOV3 cells in the trastuzumab group and the oxaliplatin group was increased(P>0.05),which was significantly increased in the combination group(P<0.05).Compared with the trastuzumab group,the S phase distribution ratio of SKOV3 cells was increased in the combination group(P<0.05).Compared with the oxaliplatin group,the S phase distribution ratio of SKOV3 cells was increased in the combination group(P>0.05).(3)Compared with the control group,the G2/M phase distribution ratio of SKOV3 cells was increased in the trastuzumab group and the oxaliplatin group(P>0.05),which was significantly increased in the combination group(P<0.05).Compared with the trastuzumab group,the G2/M phase distribution ratio of SKOV3 cells was increased in the combination group(P>0.05).Compared with the oxaliplatin group,the S phase distribution ratio of SKOV3 cells was increased in the combination group(P>0.05).3.Western blot analysis showed that:(1)the expression of p-AKT:compared with the control group,the expression of p-AKT was weakened in the trastuzumab group(P>0.05),in the oxaliplatin group(P<0.05),which was significantly weakened in the combination group(P<0.01).The expression of p-Akt was decreased in the combination group compared to that in the trast uzumab group(P>0.01).In addition,the expression p-AKT was decreased in the combination group in comparison to the oxaliplatin group(P<0.05).(2)the expression of p-ERK:compared with the control group,the expression of p-ERK was decreased in the trastuzumab group(P>0.05),in the oxaliplatin group(P<0.01),which was significantly decreased in the combination group(P<0.01).The expression of p-Erk was reduced in the combination group compared with the trastuzumab group(P<0.01),which was also decreased compared with the oxaliplatin group(P<0.05).(3)the expression of Bcl-2:compared with the control group,the expression of Bcl-2 was decreased in the trastuzumab group(P>0.05),in the oxaliplatin group(P>0.05),which was significantly decreased in the combination group(P<0.05).Compared with the trastuzumab group,the expression Bcl-2 was decreased in the combination group(P<0.05).Compared with the oxaliplatin group,the expression of Bcl-2 was reduced in the combination group(P<0.05).(4)the expression of Bax:compared with the control group,the expression of Bax was decreased in the trastuzumab group(P>0.05),increased in the oxaliplatin group(P>0.05),significantly enhanced in the combination group(P<0.05).Compared with the trastuzumab group,the expression of Bax was enhanced in the combination group(P<0.05).Compared with the oxaliplatin group,the expression of Bax was increased in the combination group(P>0.05).Conclusions:1.Trastuzumab targets HER2 protein.Oxaliplatin inhibits cell cycle distribution and inhibits proliferation of ovarian cancer SKOV3 cells by inhibiting PI3K/AKT signaling pathway and Ras/Raf/MEK/ERK signaling pathway.2.Trastuzumab combined with oxaliplatin has a synergistic effect in the treatment of HER2-positive ovarian cancer. |
