The Role Of Apatinib Combined With Aluminum-Paclitaxel In Drug-resistant Ovarian Cancer

Objective:To investigate the anti-tumor effect of Apatinib combined with different doses of albumin paclitaxel in drug-resistant ovarian cancer cell lines and xenotransplantation tumor models and the adverse reactions.Methods:1.The effect of albumin paclitaxel and Apatinib in SKOV-3/DDPSKOV-3/DDP were divided into three groups,a):Control group,no drugintervention;b):nab-P group,albumin paclitaxel 40?mol/l;c):AP group,Apatinib 50?mol/l(according to the MTT results).The expression of apoptosis related protein(Bax,BCL-2),vascular related protein(p-VEGFR-2),invasion related protein(MMP-2)were detected by Western blot and immunofluorescence,and using Transwell test to detect the invasion and migration ability of tumor cells.2.The combination effect of albumin paclitaxel and Apatinib in SKOV-3/DDPTo establish different doses of albumin paclitaxel combined with Apatinib experimental group,a):Control group,no drug intervention;b):Group-1,paclitaxel 5?mol/l+Apatinib10?mol/l,c):Group-2,paclitaxel 4.5?mol/l+Apatinib 10?mol/l,d):Group-3,paclitaxel 4?mol/l+Apatinib 10?mol/l.Using Western blot,immunofluorescence,Transwell test detect the inhibition effect.3.The combination effect of albumin paclitaxel and Apatinib in xenograft tumor modelThe results of animal experiments were compared with those of cell experiments.BALB/c female ovarian cancer nude mice were randomly divided into four groups:a)Control group,without drug intervention;b)albumin paclitaxel 20mg/kg+Apatinib150mg/kg;c):albumin paclitaxel 18mg/kg+Apatinib 150mg/kg;d):albumin paclitaxel 16mg/kg+Apatinib 150mg/kg.The anti-tumor effect was analyzed by the tumor growth curve.And the expression of Bax,BCL-2,CD31,p-VEGFR-2 and MMP-2 were detected by Western blot,immunofluorescence and immunohistochemistry.On the other hand,the drug's side effects were evaluated by recording the average daily food intake and one hour activity.Results:1.The effect of albumin paclitaxel and Apatinib in SKOV-3/DDPThe IC-50 values of albumin paclitaxel and Apatinib to SKOV-3/DDP were 45.53±4.06?mol/l(24h),20.88±2.99?mol/l(48h),8.77±0.64?mol/l(72h)and 65.74±8.33?mol/l(24h),50.66±4.96?mol/l(48h),39.547±6.62?mol/l(72h),respectively.Western blot was used to detect the relative gray values of related proteins.The values of albumin paclitaxel group and apatinib group were 0.13±0.07,0.14±0.09 (Bax),0.09±0.01,0.09±0.01(BCL-2),0.13±0.8,0.04±0.01(p-VEGFR-2),0.04±0.01,0.05±0.01(MMP-2),respectively.The positive rates of immunofluorescence were 13.87%±1.91,14.17%±1.69(Bax),0.09%±0.01,0.09%±0.01(BCL-2),25.81%±2.33,7.4%±0.88(p-VEGFR-2),10.4%±0.85,10.8%3±1.92(MMP-2),respectively.The average number of invasive cells in 24 hours was detected by Transwell test,which was 41±7.6 and 53±4.7.The difference between the experimental group and the control were all statistically significant(P<0.01).2.The combination effect of albumin paclitaxel and Apatinib in SKOV-3/DDPThe combined inhibition index CL<1 of albumin paclitaxel and Apatinib on SKOV-3/DDP,the relative gray values of combination group-1,group-2 and group-3were 0.15±0.01,0.14±0.01,0.14±0.01(Bax),0.06±0.01,0.06±0.01,0.06±0.01 (BCL-2),0.03±0.00,0.04±0.00,0.04±0.00(p-VEGFR-2),0.03±0.00,0.04±0.00,0.04±0.00(MMP-2).The positive rates of immunofluorescence were 23.07%±1.13,21.57%±0.67,21.8%±1.8(Bax),7.57%±1.11,7.4%±1.1,8.2%±1.2(BCL-2),6.8%±1.51,7.5%±0.94,8.4%±0.88(p-VEGFR-2),7.97%±1.2,9.83%±1.01,11.13%±0.4(MMP-2).The average number of invading cells of Transwell were 6,12,14.Compared with the Control group,the difference were statistically significant(P<0.01),there were no significant discrepancy between the experiment groups.3.The combination effect of albumin paclitaxel and Apatinib in xenograft modelThe average volume of tumor mass in Control group,combination group-1,group-2 and group-3 were 1452.83±61.47mm~3,144±37.65mm~3,142.33±31.4mm~3 and138.83±31.43mm~3,respectively.The discrepancy between the experimental and the control were statistically effective(P<0.01).The results of Western blot of Group-1,group-2,group-3 were as follows:0.18±0.12,0.17±0.17,0.17±0.02(Bax),0.07±0.01,0.09±0.01,0.09±0.01(BCL-2),0.16±0.02,0.16±0.04,0.2±0.01(CD31),0.16±0.03, 0.18±0.01,0.19±0.01(p-VEGFR-2),0.29±0.03,0.3±0.02,0.29±0.01(MMP-2).The positive rates of tissue protein of immunofluorescence were 20.97%±2.09,19.13%±2.78,19.73%±1.52(Bax),3.23%±1.35,2.43%±0.79,2.57%±0.5(BCL-2),6.73%±1.47,9.53%±1.6,9.7%±1.96(CD31),3.0%±0.29,3.83%±0.61,4.07%±1.04(p-VEGFR-2),2.77%±0.54,2.87%±0.49,2.93%±0.69(MMP-2).In Immunohistochemistry,the results were 34.17%±1.11,32.83%±0.95,32.77%±1.16(Bax),13.33%±1.59,14.63%±1.48,13.87%±1.89(BCL-2),11.63%±1.2,12.67%±1.52,12.33%±2.24(CD31),17.1%±1.31,17.53%±1.68,18.23%±1.25(p-VEGFR-2),14.53%±1.17,15.27%±0.74,15.57%±2.75(MMP-2).There were significant difference in the expression of related proteins between the combined groups and the control(P<0.01),and there were no obvious discrepancy between the experimental groups.Conclusions:1.Albumin paclitaxel and Apatinib all can play anti-tumor role in drug-resistant ovarian cancer cells;2.Apatinib combined with albumin paclitaxel can play a synergistic role in drug-resistant ovarian cancer cells and improve the effect of single drug;3.In combination with Apatinib,the dose of albumin paclitaxel can be reduced,and then the adverse reactions of chemotherapy drugs to the body can be reduced.