摘要
目的:基于VEGF/VEGFR2信号通路探讨中药复方肠复康(CBG)胶囊干预结肠癌血管生成的作用机制。方法:选取人结肠癌细胞HT29作为研究对象,采用低、中、高剂量CBG处理(CBG浓度分别为0.4、0.8、1.6 mmol/L),PBS处理作为溶剂对照组,显微镜下观察各组细胞形态学改变,MTT实验、Transwell小室实验、小管形成实验及ELISA试剂盒法分别检测各组细胞增殖能力、侵袭能力、血管生成能力及VEGF、VEGFR2的表达。构建人结肠癌移植瘤裸鼠模型,实验小鼠随机分为4组,每组12只:CBG低剂量组、CBG中剂量组和CBG高剂量组分别灌胃4.5、9、18 g/kg的CBG,灌胃剂量0.3 ml/d,模型组以等量生理盐水灌胃。实验28 d后杀死小鼠,比较各组小鼠的瘤体平均质量,免疫组化和Western blot法测定各组小鼠肿瘤组织血管密度(MVD)和VEGF、VEGFR2蛋白的表达。结果:经过CBG作用后HT29细胞随CBG浓度的增加发生明显的形态改变;与溶剂对照组比较,低、中、高剂量CBG处理的HT29细胞的增殖、侵袭能力和血管生成能力明显受到抑制(P<0.05),VEGF、VEGFR2的表达明显下降(P<0.05);与模型组相比,CBG低、中、高剂量组小鼠肿瘤平均质量明显降低(P<0.05),肿瘤组织MVD明显减少(P<0.05),肿瘤组织中VEGF、VEGFR2蛋白的表达水平明显下降(P<0.05),随着CBG浓度增加,体内及体外实验结果均具有剂量依赖性。结论:中药复方肠复康胶囊通过VEGF/VEGFR2信号通路干预结肠癌血管生成。
Objective:To investigate the mechanism of Chinese herbal compound Cerebiogen(CBG)capsules intervening the angiogenesis in colon cancer based on VEGF/VEGFR2 signaling pathway.Methods:The research object was human colon cancer cell line HT29,HT29 cells were treated with low,medium and high doses of CBG(CBG concentrations were 0.4 mmol/L,0.8 mmol/L,1.6 mmol/L,respectively),PBS treatment was used as the solvent control group.The morphological changes of each group were observed under microscope.MTT assay,Transwell chamber assay,tubule formation assay and ELISA kit were used to detect the cell proliferation,invasion and angiogenesis of VEGF and VEGFR2.A nude mouse colon cancer xenograft model was constructed.The experimental mice were randomly divided into 4 groups,12 in each group:model group,0.3 ml normal saline;CBG low dose group,4.5 g/kg;CBG medium dose group,9 g/kg;CBG high dose group 18 g/kg;the sham group was given 0.3 ml of normal saline.After 28 days of experimentation,the mice were killed and the tumor was observed to compare the average mass of the tumors of each group of mice.The microvessel density(MVD)of tumor tissues was detected by immunohistochemistry.The expression of VEGF and VEGFR2 protein in tumor tissues of each group was detected by Western blot.Results:After CBG treatment,HT29 cells showed obvious morphological changes with increasing CBG concentration.Compared with the solvent control group,the proliferation,invasion ability and angiogenic ability of low,medium and high dose CBG-treated HT29 cells were significantly inhibited(P<0.05).The expression of VEGF and VEGFR2 were significantly decre-ased(P<0.05).Compared with the model group,the average tumor mass of CBG low,medium and high dose groups was significantly decreased(P<0.05),and the tumor tissue MVD was significantly decreased(P<0.05),the expression levels of VEGF and VEGFR2 protein in tumor tissues were significantly decreased(P<0.05).With the increase of CBG concentration,the results in vivo and in vitro were dose-dependent.Conclusion:Cerebiogen can interfere with colon cancer angiogenesis through VEGF/VEGFR2 signaling pathway.
作者
夏雨
李涛
孙名扬
XIA Yu;LI Tao;SUN Ming-Yang(Peking University People′s Hospital,Beijing 100000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第11期1318-1323,共6页
Chinese Journal of Immunology
