健脾疏肝抗毒方对三阴性乳腺癌荷瘤鼠抗肿瘤作用的研究

The anti- tumor effect of Jian Pi Shu Gan Kang Du decoction on the triple- negative breast cancer( TNBC) tumor- bearing mice

目的研究健脾疏肝抗毒方对三阴性乳腺癌荷瘤鼠抗肿瘤作用及其可能的机制。方法建立稳定高表达的p Ac GFP MDA-MB-231细胞株,采用皮下接种法建立MDA-MB-231-p Ac GFP细胞异种移植瘤裸鼠,应用免疫组化法检测不同组别:对照组(A组)、健脾疏肝抗毒方组(B组)、ADM3100(CXCR4抑制剂)组(C组)、健脾疏肝抗毒方联合ADM3100组(D组)肿瘤微环境中的肿瘤相关巨噬细胞、肿瘤相关成纤维细胞浸润,蛋白印迹方法检测各组肿瘤组织CXCR4/CXCL12轴相关分子的表达。结果健脾疏肝抗毒方能显著抑制肿瘤微环境中肿瘤相关巨噬细胞、肿瘤相关成纤维细胞的浸润,抑制CXCR4/CXCL12轴相关分子CXCR4、CXCL12、HIFα,NF-κB,VEGF的表达,且与ADM3100联合后抑制作用显著增强。结论健脾疏肝抗毒方可以通过改变肿瘤微环境,调节CXCR4/CXCL12轴相关分子的表达而发挥抑制乳腺癌荷瘤鼠异种移植瘤的生长的作用。

Objective To investigate the anti - tumor effectof JianPi ShuGan Kang Du decoction on the triple - negative breast cancer（ TNBC ） tumor - bearing mice and study its possible ,nechanism . Methods Construct stable highexpression cell lines （pAcGFP MDA- MB- 231 ） , establish the xenograft model of human breast cancer in BALB/c mice by subcutaneous inoculation. Examine theinfihration of tumor associated macrophages （TAMs） and cancer -associated fibroblasts （CAFs） in tumour microenvironmen in differrnt groups with Immunohisochemical staining. Using Western Blot to test the expression of association protein in different gruops（ including CXCR4, CXCL12, HIFα, NF - κB, VEGF）. Results JianPi ShuGan KangDu decoction can significantly suppress the infiltration of tumor associated macrophages （ TAMs ） and cancer - associated fibroblasts （ CAFs ）, and ADM3100 can enhance that effection. Protein expression of group B, C and D are inhibited with varying degrees, also we found group D has a higher inhibition ratio. Conclusion JianPi ShuGan KangDu decoction can regulate CXCR4 / CXCL12 axis related molecules expression through changing the tumor microenvironment, so that it could inhibit the growth and liver metastasis of human breast cancer xenograft in nude micer.