健脾消癌方对结肠癌肝转移裸鼠模型肿瘤微环境转移相关因子表达的影响

Effect of Jianpi Xiaoai Prescription on Expression of Metastasis-related Factors in Tumor Microenvironment of Nude Mouse Model of Liver Metastasis of Colon Cancer

目的:研究健脾消癌方对结肠癌肝转移裸鼠模型肝组织中趋化因子受体4(CXCR4)、转化生长因子-β(TGF-β)、整合素αvβ5(ITGαvβ5)、血清钙结合蛋白A4(S100A4)、血清钙结合蛋白A8(S100A8)、血清钙结合蛋白A9(S100A9)等肿瘤微环境转移相关因子蛋白表达的影响,探讨其抗结肠癌肝转移的可能作用机制。方法:将BALB/c裸鼠随机分为正常组、模型组、健脾消癌方低、中、高剂量组。建立人结肠癌肝转移裸鼠模型。健脾消癌方低、中、高剂量组分别灌胃5.4、10.8、21.6 g·kg^(-1)药液,模型组及正常组给予等体积蒸馏水灌胃,每天给药1次,连续给药3周。末次给药后24 h脱颈处死,观察各组裸鼠肝转移情况,并通过蛋白免疫印迹法(Western blot)检测肝组织中CXCR4、TGF-β、ITGαvβ5、S100A4、S100A8、S100A9等肿瘤微环境中转移相关因子蛋白表达情况。结果:模型组、健脾消癌方低、中、高剂量组转移瘤面积占比分别为73%、72%、55%、42%。健脾消癌方高剂量组明显低于模型组(P<0.05)。与模型组比较,健脾消癌方高、中剂量组CXCR4、TFG-β、ITGαvβ5、S100A8、S100A9表达均明显降低(P<0.05);健脾消癌方高、中、低剂量组S100A4表达均明显降低(P<0.05)。健脾消癌方低剂量组CXCR4、TFG-β、ITGαvβ5、S100A8、S100A9与模型组比较表达差异无统计学意义。结论:健脾消癌方能抑制结肠癌肝转移,其机制可能与降低CXCR4、TGF-β、ITGαvβ5、S100A4、S100A8、S100A9等肿瘤微环境中转移相关因子的表达相关。

Objective:To study the effect of Jianpi Xiaoai prescription on the protein expression of metastasis-related factors in tumor microenvironment such as chemokine receptor 4(CXCR4),transformation growth factor-β(TGF-β),integrin avβ5(ITGαvβ5),serum calcium-binding protein A4(S100A4),serum calcium-binding protein A8(S100A8),and serum calcium-binding protein A9(S100A9)in the liver tissue of the nude mouse model of liver metastasis of colon cancer,and to explore the possible mechanism of its anti-liver metastasis of colon cancer.Method:BALB/c nude mice were randomly divided into blank group,model group,and Jianpi Xiaoai prescription low,medium,and high-dose groups.A nude mouse model of liver metastasis of human colon cancer was established.Jianpi Xiaoai prescription low,medium,and high-dose groups were given 5.4,10.8,21.6 g·kg^(-1)liquid medicine,respectively,and the model group and the blank group were given the same volume of distilled water by gavage,once a day for 3 consecutive weeks.24 h after the last administration,the nude mice were sacrificed by neck removal,and the liver metastasis of each group was observed.Western blot was used to determine the protein expression of metastasis-related factors in the tumor microenvironment,such as CXCR4,TGF-β,ITGαvβ5,S100A4,S100A8,and S100A9.Result:The proportion of metastatic tumor area was 73%in the model group,72%in the low-dose group,55%in the medium-dose group,and 42%in the high-dose group.The high-dose group was significantly lower than the model group(P<0.05).As compared with the model group,the expression of CXCR4,TFG-β,ITGαvβ5,S100A8,and S100A9 in the Jianpi Xiaoai prescription high and medium-dose groups were significantly decreased(P<0.05).As compared with the model group,the expression of S100A4 in the Jianpi Xiaoai prescription high,medium,and low-dose groups was significantly decreased(P<0.05).The expression of CXCR4,TFG-β,ITGαvβ5,S100A8,and S100A9 in the Jianpi Xiaoai prescription low-dose group was not significantly different from that in the model group.Conclusion:Jianpi Xiaoai prescription can inhibit liver metastasis of colon cancer,and its mechanism may be related to the reduction of the expression of metastasisrelated factors such as CXCR4,TGF-β,ITGαvβ5,S100A4,S100A8,and S100A9 in tumor microenvironment.