防己诺林碱对结肠癌细胞恶性生物学行为及HIF-1α/VEGF/Akt通路的相关性研究  

Study on malignant biological behavior and HIF-1α/VEGF/Akt pathway of colon cancer cells by using Fangchinoline

作　　者：望永鼎[1] 刘文华[1] 翟一飞 张涛[1] WANG Yong-Ding;LIU Wen-Hua;ZHAI Yi-Fei;ZHANG Tao(Department of Surgical,Shangqiu Medical College,Shangqiu 476000,China)

机构地区：[1]商丘医学高等专科学校外科,商丘476000 [2]河南省肿瘤医院,郑州450008

出　　处：《中国免疫学杂志》2019年第24期3042-3047,共6页Chinese Journal of Immunology

基　　金：河南省教育厅高等学校重点科研项目(No.19B320013)资助

摘　　要：目的:研究防己诺林碱(FAN)对结肠癌的作用以及与缺氧诱导因子1α(HIF-1α)/血管内皮生长因子(VEGF)/蛋白激酶B(Akt)通路的相关性。方法:用5、10、20μg/ml的防己诺林碱分别处理结肠癌细胞HT-29 48 h,克隆形成实验检测细胞生长,蛋白印迹检测增殖标记蛋白Ki67、增殖细胞核抗原(PCNA)、上皮细胞钙黏蛋白(E-cadherin)、神经细胞钙黏蛋白(N-cadherin)、HIF-1α、VEGF和Akt的表达,Transwell检测细胞侵袭情况,显微镜观察上皮向间质的形态转变;采用皮下注射结肠癌细胞HT-29构建移植瘤模型,腹腔分别注射5、10、20μg/ml防己诺林碱,检测结肠癌移植瘤体积,免疫组化检测Ki67和VEGF,蛋白印迹检测HIF-1α、VEGF、Akt的表达,HE染色观察肝肾损伤。结果:与Blank cells相比较,FAN组的结肠癌细胞HT-29克隆形成率显著降低,Ki67和PCNA的表达量显著下调,侵袭细胞数目显著减少,VEGF和N-cadherin表达量显著下调,E-cadherin表达量显著上调,HIF-1α、VEGF和Akt表达量显著下调;与Blank组相比,FAN组的移植瘤体积明显小,Ki67和VEGF阳性比率显著降低,HIF-1α、VEGF和Akt表达量显著下调,肝肾无损伤。结论:防己诺林碱都可以抑制结肠癌细胞HT-29的生长和增殖、侵袭能力、上皮间质转化,并且抑制结肠癌细胞HIF-1α/VEGF/Akt信号通路,从而抑制结肠癌的发生和发展。Objective:To investigate the effects of Fangchinoline on colon cancer and its association with the HIF-1α/VEGF/Akt pathway.Methods:Colon cancer cells HT-29 were treated with 5,10,20 μg/ml Fangchinoline in vitro for 48 h.Cell growth was detected by colony formation assay.Proliferation marker proteins Ki67,PCNA,E-cadherin,N-cadherin,HIF-1α,VEGF,Akt were detected by Western blot.Cell invasion was detected by TRANSWELL,Epithelial-to-mesenchymal morphological transformation was observed by microscopic.The transplanted tumor model was constructed by subcutaneous injection of colon cancer cell line HT-29,and 5,10,20 μg/ml anti-ennoline was injected into the abdominal cavity.The volume of colon cancer xenografts was detected,Ki67 and VEGF were detected by immunohistochemistry,and the expressions of HIF-1α,VEGF and Akt were detected by Western blot,liver and kidney injury was observed by HE staining.Results:Compared with Blank cells,the colony of HT-29 colon cancer cells in FAN group was significantly decreased,the expression levels of Ki67 and PCNA were significantly down-regulated,the number of invasive cells was significantly decreased,and the expression levels of VEGF and N-cadherin were significantly down-regulated.The expression of E-cadherin was significantly up-regulated,and the expression levels of HIF-1α,VEGF and Akt were significantly down-regulated.Compared with the Blank cells group,the tumor volume of the FAN group was significantly smaller,and the positive ratio of Ki67 and VEGF was significantly decreased.The expression levels of HIF-1α,VEGF and Akt were significantly down-regulated.But liver and kidney were not damaged.Conclusion:Fangchinoline can inhibit the growth and proliferation of colon cancer cells HT-29,invasion,epithelial-mesenchymal transition,and inhibit the HIF-1α/VEGF/Akt signaling pathway in colon cancer cells,thereby inhibiting the occurrence and development of colon cancer.

关 键 词：防己诺林碱 结肠癌细胞HT-29 HIF-1Α VEGF Akt 

分 类 号：R734.2[医药卫生—肿瘤]