Inhibitory Effect Of Inositol Hexaphosphate And Inositol Combined With Capecitabine On The Growth And Liver Metastasis Of Colorectal Cancer In BALB/c Mice

Objective:An orthotopic transplantation model of colorectal cancer in mice was established to study the effect and mechanism of combined use of phytic acid(IP6)and inositol(INS)to assist capecitabine in inhibiting colorectal cancer growth and liver metastasis.Methods:The orthotopic implantation model of colorectal cancer in BALB/c male mice was established by injecting 0.2 m L 1×106 CT-26 cells under the serosa layer of the cecal wall of male mice.The tumor growth status of model mice was monitored by in vivo imaging technique of small animals.After 3 weeks,the tumor-forming mice were divided into 4 groups according to the cross combination of IP6+INS and capecitabine and the different levels of the two factors,divided into model group,capecitabine(60 mg/kg)group,IP6+INS(80 mg/kg:80 mg/kg)group and capecitabine + IP6+INS(60 mg/kg:80 mg/kg:80 mg/kg)group.Mice in each experimental group were intragastrically fed with the corresponding intervention 0.2 m L,and mice in the model group were fed with the same dose of normal saline for 5 days a week for 6 weeks.Observe and record the body weight,diet,drinking water and survival of mice.According to the results of in vivo imaging of small animals,record the number of tumor photons developed,calculate the tumor area.After the mice were killed,the tumor tissue in situ was weighed,and the concentrations of TNF-α,IL-6,IL-8,CCL20,IL-10 and IL-12 in serum were detected by ELISA.RT-PCR and Western blot methods were used to detect the expression of E-cadherin,N-cadherin,Vimentin MMP-2,MMP-9 m RNA and protein in mouse tissues.Results: 1.After grouping,the average body weight of the mice in each group showed a gradual upward trend.After 3 weeks of intervention,the average body weight of the mice in the IP6+INS group and the capecitabine group were higher than the model group mice,but lower than the capecitabine+IP6+ INS group;from the 5th week to the end of the experiment,except for the capecitabine+IP6+INS group,the average weight of the mice in each group showed a downward trend,and the average weight of the model group was the most obvious.The daily intake and drinking water of the mice in each group were higher than those of the model group,of which the capecitabine+IP6+INS group had the highest intake and drinking water.The survival time of mice in the capecitabine+IP6+INS group was significantly higher than that in the model group(χ2=13.14,P<0.05).2.Factorial analysis of variance analysis showed that the weight of in situ tumors in each intervention group was significantly lower than that in the model group(F=13.772-157.613,P<0.05),capecitabine + IP6+INS had the most significant effect on reducing the tumor weight in situ in mice,and there was a synergistic effect between capecitabine and IP6+INS.(F=13.772,P<0.05).3.The liver metastasis rate of each intervention group was lower than that of the model group(33.33%).The capecitabine+IP6+INS group had the lowest liver metastasis rate,reaching 8.3%,but the difference was not statistically significant(P>0.05).The analysis of variance analysis of factorial design showed that the tumor development area of mice in each intervention group was significantly lower than that in the model group(F=5.980-80.097,P<0.05).The intervention of capecitabine+IP6+INS reduced the tumor development area of mice.The effect is the most significant.Capecitabine and IP6+INS have a synergistic effect(F=5.980,P<0.05).4.Factorial design analysis of variance showed that the levels of TNF-α,IL-6,CCL20,IL-8,and IL-10 in the intervention groups were significantly lower than those in the model group(F=5.731-1013.425,P<0.05),IL-12 expression level was significantly higher than the model group(F=25.189-165.417,P<0.05),capecitabine+IP6+INS intervention reduced the expression of TNF-α,IL-6,CCL20,IL-8,IL-10 The level and the effect of increasing the expression level of IL-12 are the most significant.IP6+INS has a synergistic effect with capecitabine(F=5.731-70.491,P<0.05).5.Factorial design analysis of variance results showed that the levels of N-cadherin,Vimentin,MMP-2,MMP-9 m RNA and protein expression in the intervention groups were significantly lower than those in the model group(F=5.348-271.893,P<0.05),The E-cadherin m RNA and protein expression levels were significantly higher than the model group(F=5.348-129.552,P<0.05),capecitabine+IP6+INS decreased the m RNA and protein expression of N-cadherin,Vimentin,MMP-2,MMP-9 Level,the effect of increasing E-cadherin m RNA and protein expression level is the most significant.Capecitabine and IP6+INS have a synergistic effect(F=5.348-47.498,P<0.05).Conclusion:The combined use of IP6 and INS can effectively assist capecitabine in inhibiting the growth of colorectal cancer and the occurrence of liver metastasis.It can also improve the quality of life of tumor mice and increase the survival time of tumor mice.Its mechanism of action is related to affecting the expression of inflammatory factors,inhibiting the expression of matrix metalloproteinases,increasing the expression of epithelial markers,and inhibiting the expression of mesenchymal markers,thereby inhibiting the process of epithelial-mesenchymal transition.