Effects of Xiangcao Fuzheng Mixture on tumor invasion and metastasis and NF-κB/p65,NSD2,PD-L2 protein expression in colon cancer mice
Objective:To investigate the effect of Xiangcao Fuzheng Mixture(XCFZ)on immunosuppression and the expression of nuclear factor-kappa B(NF-κB)/p65,nuclear receptor binding SET domain-protein 2(NSD2)and programmed cell death-ligand 2(PD-L2)in colon cancer tumor-bearing mice.Methods:HT29 colon cancer cell line was cultured and transplanted to establish colon cancer tumor-bearing mice models.The mice were divided in-to several groups,including a model group,a 5-fluorouracil(5-FU)group,and XCFZ groups with low,medium,and high doses,as well as a combination group,with 10 mice per group.From the second day of modeling,the Chinese medicine group was treated with low,medium and high doses of XCFZ by gavage,the chemotherapy group was treated with intraperitoneal injection of 5-FU,the combination group was treated with medium dose of XCFZ combined with intraperitoneal injection of 5-FU,and the model group was injected with the same amount of normal saline.The body weight,tumor weight and body weight after tumor removal were measured,and the tumor inhibition rate and spleen and thymus index were calculated.The changes of CD3+.CD4+and CD8+cells were detected by flow cytometry,and the CD4+/CD8+ratio was calculated.The expression levels of CXCR4,ITGαVβ5,TGF-β and IL-10 in serum of mice were detected by ELISA.The protein levels of NF-κB/p65,NSD2 and PD-L2 in tumor tissues were detected by Western blot.Results:Compared with the model group,the com-bined group showed the most significant tumor inhibition rate,followed by the 5-FU group and the medium dose XCFZ group(P＜0.01).Compared with 5-FU group,the body weight of XCFZ low,medium,and high dose groups and XCFZ combined group increased after tumor removal(P＜0.05).Compared with 5-FU group,the spleen index of XCFZ low,medium and high dose groups and XCFZ combined group were increased(P＜0.05,P＜0.01).Compared with the XCFZ medium dose group,the spleen and thymus indexes of the XCFZ low and high dose groups,5-FU group and combination group were decreased(P＜0.05,P＜0.01).Karyopyknosis,cell necrosis and nuclear fragmentation were more common in XCFZ group.In the 5-FU group,the tumor cells were smaller,the mitotic phenomenon and atypical cells were less,and the necrotic and fragmented cells were more.Compared with the XCFZ medium dose group,CD8+cells in the XCFZ high and low dose groups,5-FU group and combination group were significantly decreased(P＜0.01).Compared with 5-FU group,CD4+/CD8+in XCFZ high,medium,low dose groups and combination group were significantly decreased(P＜0.05,P＜0.01),and the ratio in XCFZ medium dose group was the lowest(P＜0.05,P＜0.01).Compared with the 5-FU group,the expression levels of CXCR4,ITGαVβ5,TGF-β and IL-10 in the combined group were significantly decreased(P＜0.05,P＜0.01).The expression levels of CXCR4,ITGαVβ5,TGF-β and IL-10 in the high and low dose XCFZ groups were significantly higher than those in the medium dose XCFZ group(P＜0.05,P＜0.01).Compared with the medium dose XCFZ group,the high and low dose XCFZ groups had significant increases in the expression of NF-κB/p65,NSD2 and PD-L2(P＜0.05,P＜0.01).Compared with the 5-FU group,the combined group had a better inhibitory effect on the expression of NF-κB/p65,NSD2 and PD-L2(P＜0.01).Conclusion:XCFZ can in-hibit the tumor growth of colon cancer bearing mice,and the medium dose has the best effect.XCFZ can reduce the serum level of tumor metastasis factors,improve the immunosuppressive state,and down-regulate the expres-sion of NF-κB/p65,NSD2,and PD-L2 related proteins,thereby inhibiting the invasion and metastasis of colon cancer.
NETL
NSTL
万方数据
