Effect Of IP6 + Ins On The Expression Of VEGF And IGF1 And Their Invasion Ability In Human Ovarian Cancer SKVO3 Cell Line

Ovarian cancer is one of the most common gynecological malignancies.It spreads extensively in the pelvis and abdomen,and its mortality is the highest in gynecologic malignancies.The 5-year survival rate is only about30%^[1].Although most ovarian cancer patients initially benefit from satisfactorycytoreductivesurgerycombinedwithplatinum-based chemotherapy,however,nearly 90%of patients still experience relapse^[2],develop resistance and eventually die.Like most malignant solid tumors,neoplasms that are rich in neovascularization promote the growth,invasion and metastasis of ovarian cancer,with poor prognosis^[3].VEGF（vascular endothelial growth factor）and IGF-1（insulin-like growth factor-1）are overexpressed in many types of tumors,including ovarian cancer,both of which promote neovascularization and invasion of ovarian cancer^[4].Although targeted drugs targeting VEGF and IGF-1 have been used clinically,they are not only expensive but also produce many side effects with poor clinical outcomes.Faced with such a severe situation,looking for a new treatment to improve the clinical efficacy of ovarian cancer is particularly important.IP6,also known as phytate or phytic acid,is safe and non-toxic and can be found in almost all mammals and plants^[5].IP6 has a wide range of anti-cancer effects,showed inhibition of many types of tumors in a dose-dependent manner.In addition,IP6 in combination with other drugs can enhance the anti-cancer effect,while reducing the side effects of anti-cancer drugs.Ins,which is called inositol,also has moderate antitumor activity.The combination of IP6+Ins has a coordinating effect,and the anticancer effect is obviously superior to any one used alone^[6].Objective:The purpose of this study was to investigate the effects of different doses of IP6+Ins,the combination of IP6+Ins and cisplatin on the expression of VEGF and IGF-1 in ovarian cancer cell line SKOV3 and their effect on tumor cell invasion,hoping to provide new ideas for the treatment of ovarian cancer.Methods:1.Cell Culture:Human ovarian cancer SKVO3 cell lines were cultured using conventional cell culture techniques.2.Experimental grouping:The experiment was divided into 6 groups:a,b,c,d,e and f,respectively,in which“a”was the blank control group,“b”was the IP6+Ins low dose group（1.5 mmol/L）,“c”was the IP6+Ins medium dose group（3mmol/L）,“d”was the IP6+Ins high dose group（6mmol/L）,“e”,for the cisplatin group（3ug/ml）,“f”,for the combination group:cisplatin+（IP6+Ins）medium dose group（3 ug/ml+3 mmol/L）.3.Transwell cell invasion assay observed the different concentrations of IP6+Ins,cisplatin and the combination of both on the invasion of human ovarian cancer SKOV3 cells.4.The effects of cisplatin,different concentrations of IP6+Ins and combined treatment on the expression of VEGFmRNA and IGF-1mRNA in SKOV3 human ovarian cancer cells were detected by RT-PCR.5.Western blot was used to detect the expression of IGF-1 in human ovarian cancer cell line SKOV3.6.Statistical methods:The experiment use SPSS22.0 statistical software to process and analyse the experimental data,P<0.05 indicated the difference was statistically significant.Results:1.The results of Transwell cell invasion assay showed that the cell numbers of a,b,c,d,e and f in each group was:77.67±4.16,65.00±3.61,50.33±4.51,39.00±3.00,24.33±3.06,16.67±1.53,respectively.Compared with the control group,the number of cells in each group decreased,the difference was statistically significant（P<0.05）.At the same time,the number of cells in different concentrations of IP6+Ins group was ranked as:b group>c group>d group,and the differences were statistically significant（P<0.05）.In addition,the order of cell numbers in IP6+Ins middle dose group,cisplatin group and combination group was as follows:c group>e group>f group,with significant difference after any comparison（P<0.05）.2.The expression of VEGF mRNA in SKOV3 cells was detected by RT-PCR.VEGF mRNA was found in all the groups,which was high in the control group and low in the treatment group.In all the groups,the expression of VEGF mRNA was as follows:a group>b group>c group>d group>e group>f group,and the differences were statistically significant（P<0.05）.3.IGF-1mRNA expression in each group:the expression of IGF-1mRNA in a,b,c,d group was:1.000±0.000,0.8900±0.0139,0.8014±0.0307,0.7187±0.0179,respectively.With the increase of the concentration,the expression of IGF-1 mRNA gradually decreased.The expression of IGF-1mRNA in c,e and f groups was 0.8014±0.0307,0.5960±0.0254 and 0.4985±0.156,respectively,which means c group>e group>f group.The differences among the groups were statistically significant（P<0.05）.4.Western blot test detected the expression of IGF-1 protein in SKOV3human ovarian cancer cells.The order of the expression of IGF-1 protein in all the groups from high to low was:a group>b group>c group>d group>e group>f group,and the difference was statistically significant（P<0.05）.Conclusions:1.IP6+Ins can reduce the expression of VEGF mRNA in ovarian cancer SKOV3 cells.2.IP6+Ins can reduce the expression of IGF-1 mRNA and IGF-1 protein in ovarian cancer SKOV3 cells.3.IP6+Ins can inhibit the invasion of ovarian cancer SKOV3 cells in a dose-dependent manner.IP6+Ins may inhibit its invasion by downregulating the expression of VEGF and IGF-1.4.The combination of IP6+Ins and cisplatin has a coordinated effect.IP6+Ins may enhance the inhibitory effect of cisplatin on human ovarian cancer SKOV3 cells by decreasing the expression of IGF-1 mRNA and IGF-1protein.