白花蛇舌草通过调控Wnt/β-catenin通路抑制大肠癌细胞及大肠癌干细胞的生长  

Hedyotis diffusa Willd inhibits the growth of colorectal cancer cells and stem cells via suppressing Wnt/β-catenin signaling pathway

作　　者：林久茂[1] 魏丽慧[1] 李琼瑜[1] 赖子君 彭军[1] 

机构地区：[1]福建中医药大学中西医结合研究院,福州350122

出　　处：《中华中医药杂志》2015年第5期1805-1808,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：博士点基金博导类(No.20133519110003)~~

摘　　要：目的:从Wnt/β-catenin信号通路调控探讨白花蛇舌草(HDW)抑制大肠癌细胞及其干细胞生长的作用机制。方法:MTT法检测HDW对HT-29细胞活力的影响;建立HT-29皮下移植瘤模型,通过测量瘤体体积观察HDW对瘤体生长的影响,采用免疫组化检测HDW干预后对移植瘤组织中ki-67表达的影响;采用SP法分析HDW对HT-29细胞干细胞比例的影响;同时,采用Western blotting及免疫组化法检测HDW干预后对HT-29细胞中干细胞标记物LGR-5、OCT-4表达和Wnt/β-catenin通路中APC、survivin、β-catenin及p-βcatenin表达的影响。结果:MTT检测结果显示不同浓度的HDW干预可显著降低HT-29细胞活力,呈现一定的剂量依赖;HDW显著抑制HT-29移植瘤瘤体的生长且能够下调ki-67的表达;HDW可降低HT-29细胞中干细胞的比例,体内外实验结果均显示HDW可明显抑制干细胞标记物LGR-5和OCT-4的表达,并且上调APC及p-β-catenin的表达,下调survivin的表达。结论:HDW对大肠癌细胞和大肠癌干细胞的生长具有显著抑制作用,抑制Wnt/β-catenin信号通路的活化及调控关键因子的表达是其重要作用机制之一。Objective： To investigate the effect of Hedyotis diffusa Willd （HDW） on growth of colorectal cancer cells （CRC） and and colorectal cancer stem cells （CRCSC）, and explore the underlying mechanism on Wnt/β -catenin pathway in vivo and in vitro. Methods： MTT assay was used to determine the cell viability of HT-29 cells with the treatment of HDW. Using a colorectal cancer mouse xenograft model, the effect of HDW on tumor growth was evaluated by measuring the tumor volume. Immunohistochemistry method was used to detect the expression of ki-67 in tumor tissue after treated by HDW. SP analysis was uesd to determine the percentage of cancer stem cells. Western blotting and immunohistochemistry were performed to determine the expression of LGRS, OCT-4 and relative genes involved in Writ/β -catenin pathway such as APC, survivin,β -catenin and p- β catenin in HT-29 cells and tumor tissue. Results： The treatment of HDW significantly decreased the cell viability and inhibited the tumor growth and expression of ki-67 of HT-29 cells, demonstrating that HDW could inhibit CRC growth in vitro and in vitro. Moreover, SP analysis indicated that HDW treatment significantly decreased the percentage of stem cells and decreased the expression of biomarkers （LGR-5 and OCT-4） of CSC in HT-29 cells in vitro and in vitro. In addition, HDW treatment increased the phosphorylation β -catenin and up-regulated the epxression of APC, and down-regulated the expression of survivin. Conclusion： These results indicate that suppression of the Wnt/β -catenin pathway might be one of the mechanisms by which HDW inhibits the growth of CRC and CRCSCs.

关 键 词：白花蛇舌草 大肠癌 WNT/Β-CATENIN信号通路 肿瘤干细胞 

分 类 号：R285[医药卫生—中药学]