In Vitro And In Vivo Exploration Of Genistein Combined With Cisplatin On Human Ovarian Carcinoma And Associated Mechanism

Ovarian cancer is one of the most frequently gynecological malignancies,which often needs adjuvant chemotherapy because of the difficullty of early diagnosis and surgery. Although 70%-80%of patients respond to platinum-based chemotherapy,the majority will relapse and ultimately succumb to metastatic disease because of the side effect of long-term chemotherapy and the emergence of chemoresistance,which made chemotherapy restricted.Furthermore,ovariotomy often needed in ovarian cancer,yong patients often consulted whether they could complement estrogen to alleviate the discomfort due to ovariotomy.Nevertheless,previous research found estrogen could promote proliferation and metastasis of ovarian cancer cell.Therefore people are searching for new remedy that could alleviate the discomfort of low estrogen,kill tumor cell,decrease side effect of chmotherapy and chemoresistance,and also have anticancer action.Genistein,a natural isoflavenoid found in soybean products,has been reported to have both chemopreventive and chemotherapeutic potential in multiple tumor types. Several in vitro and in vivo studies have documented its anti-neoplastic effects in prostate,breast,colon and pancreatic adenocarcinomas.Genistein can also inhibit the proliferation of choriocarcinoma and ovarian cancer cells.The exact pharmacological mechanism by which genistein exerts this properties and whether it has cooperation with chemotherapy has not been determined.In this study,we have investigated the effect of genistein,cisplatin,genistein combined with cisplatin on the growth inhibition of ovarian cancer SKOV₃ cells,the inhibitory effect of genistein,cisplatin,genistein combined with cisplatin on human ovarian carcinoma exnograft tumors and associated mechanism,evaluated its feasibility as a potential chemotherapeutic and chemopreventive agent in the treatment of chemoresistant ovarian cancer.There are two parts in the study:Partâ… Effects of genistein combined with cisplatin on proliferation and apoptosis in the ovarian cancer cell line SKOV₃ and its relationship with VEGF and u-PA expression.Partâ…¡Inhibitory effects of genistein combined with cisplatin on ovarian carcinoma xenograft tumors and its relationship with VEGF and u-PA expression.Partâ… Effects of genistein combined with cisplatin on proliferation and apoptosis in the ovarian cancer cell line SKOV₃ and its relationship with VEGF and u-PA expressionObjective To study the effects of genistein,cislpatin(DDP),genistein combined with DDP on morphological changes,proliferation and apoptosis of human ovarian cancer cell line SKOV₃ and its relationship with VEGF(vascular epithelial growth factor) and u-PA(urokinase type plasminogen activator) expression.Materiol and Methods Genistein and DDP was incubated in different concentrations with ovarian cancer SKOV₃.The effect of genistein on morphological changes of SKOV₃ cells were observed by optical microscope.The effect of genistein and DDP on SKOV₃ cell proliferation was measured by MTT assay.Cell apoptosis were analyzed by flow cytometry(FCM).The VEGF and u-PA mRNA expression of SKOV₃ cell were detected by RT-PCR analysis.Result 1umol/L of DDP had weak inhibitory effect on SKOV₃ cell,more than 5umol/L of DDP had remarkable inhibitory effect on SKOV₃ cell,which had statistical difference compared with control group(P＜0.05); 5umol/L of genistein had weak inhibitory effect on SKOV₃ cell,more than 10umol/L genistein had distinct inhibitory effect on SKOV₃ cell,which had statistical difference compared with control group(P＜0.05);5μmol/L of DDP combined with genistein(5,10,20μmol/L)could significantly inhibit the proliferation of ovarian cancer SKOV₃ cell,wich had statistical difference compared with single medication group (P＜0.01).Genistein,DDP used alone and in combination with each other inhibited the proliferation of ovarian cancer SKOV₃ cell in a time and dose dependent manner,both of them had cooperation effect.Genistein combined with DDP significantly induced the apoptosis of SKOV₃ cell compared with control and genistien,DDP used alone group,wich had statistical difference(P＜0.01).Genistien combined with DDP significantly lower the VEGF,u-PA mRNA expression of SKOV₃ cell compared with control and genistein,DDP used alone group,wich had statistical difference(P＜0.01).Conclusion Genistein compared with DDP can significantly inhibit proliferation,induce differentiation and apoptosis of ovarian cancer SKOV₃ cell,which is associated with the VEGF and u-PA expression.Partâ…¡Inhibitory effects of genistein combined with cisplatin on ovarian carcinoma xenograft tumors and its relationship with VEGF and u-PA expressionObjective To explore the inhibitory effect of genistein,cislpatin(DDP),genistein combined with DDP on human ovarian carcinoma exnografi tumors and its relationship with VEGF(vascular epithelial growth factor) and u-PA(urokinase type plasminogen activator) expression.Materiol and Methods Human SKOV₃ ovarian carcinoma xenograft models were established in BALB/c nude mice,and the nude mice were randomly divided into four groups:control group,DDP treated group,genistien treated group,genistein plus DDP treated group.The growth ability of xenograft tumors were observed every 2 days.The VEGF expression of xenograft tumors was analyzed by immunohistochemical staining,RT-PCR and Western blot. The u-PA expression of xenograft tumors was analyzed by RT-PCR and western blot. Result Genistein,DDP could inhibit the growth of xenograft tumors.The volume and weight of each xenograft tumors was less than control group.The inhibitory rate of DDP,genistein used alone groups and combination groups was 33.1%,43.6%,58.7%,wich had statistical difference compared with control group (P＜0.01).The VEGF,u-PA mRNA and protein expression were significantly lower in genistein combined with DDP treated xenograft tumors than genistein and DDP used alone xenograft tumors,wich had statistical difference(P＜0.01).Conclusion genistein combined with DDP can significantly inhibit the growth ability of ovarian carcinoma xenograft tumors,which is associated with the VEGF and u-PA expression.