半枝莲通过Hedgehog信号通路抑制结肠肿瘤干细胞自我更新  

Scutellaria barbata inhibits the self-renewal of colonic cancer stem cells through Hedgehog signaling pathway

作　　者：卢小路 徐海波[1] 石梦莹[1] 史玉荣[1] 周丽娟[2] 王德健[1] 陈继兰[3] 方皓[1] 谭溧 周雪[1] 

机构地区：[1]成都中医药大学药学院、中药资源系统研究与开发利用省部共建国家重点实验室培育基地,成都611137 [2]四川省中医药科学院中医研究所,成都610014 [3]成都中医药大学基础医学院,成都610037

出　　处：《中药药理与临床》2015年第1期139-141,共3页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(81173598);四川省科技厅"中药药理四川省青年基金科技创新研究团队"(2014TD0007);成都中医药大学科技发展基金(ZRYY201201)

摘　　要：目的:研究半枝莲对结肠肿瘤干细胞(CSC)自我更新的影响,从Hedgehog信号通路阐明其作用机制。方法:制备半枝莲乙醇提取物,从HT-29人结肠腺癌上皮细胞中分拣出结肠CSC,用Aqueous法,研究半枝莲对结肠CSC体外活性的影响,用干细胞球形成试验研究半枝莲对结肠CSC自我更新的影响。用qRT-PCR研究半枝莲对结肠CSC标记物基因CD133和Lgr5表达的影响,和对结肠CSC中Hedgehog信号通路基因Ptch1和Gli1表达的影响。结果:终浓度为125μg/ml、250μg/ml和500μg/ml的半枝莲乙醇提取物可以剂量依赖性地抑制结肠CSC体外活性,抑制结肠CSC自我更新,下调结肠CSC标记物基因CD133和Lgr5 mRNA水平,降低结肠CSC中Hedgehog信号通路基因Ptch1和Gli1 mRNA转录。结论:半枝莲可以抑制结肠CSC自我更新,其机制在于抑制Hedgehog信号通路的活性。Objective： To investigate the effect of Scutellaria barbata D. Don（ SBDD） on the self-renewal of colonic cancer stem cells（ CSC） and its underlying mechanism through Hedgehog signaling pathway. Methods： The ethanol extract of SBDD was prepared and the colonic CSC were fractionated from human colonic adenocarcinoma epithelial cells HT-29. The effect of SBDD on cell viability of colonic CSC was determined with Aqueous method. The function o SBDD on colonic CSC self-renewal was assessed with spheroid formation assay. The effects of SBDD on the expressions of colonic CSC marker genes including as CD133 and Lgr5,and the expressions of Ptch1 and Gli1 involved in Hedgehog signaling pathway of colonic CSC were measured with quantitative real-time RT-PCR. Results： The ethanol extract of SBDD at final concentrations of 125 μg / ml,250 μg / ml and 500 μg / ml dose-dependently inhibited colonic CSC viability and its self-renewal,down regulated mRNA level of colonic CSC marker genes including as CD133 and Lgr5,and decreased the mRNA transcription of Ptch1 and Gli1 involved in Hedgehog signaling pathway of colonic CSC. Conclusions： SBDD suppresses colonic CSC self-renewal by reducing the activity of Hedgehog signaling pathway.

关 键 词：半枝莲 肿瘤干细胞 自我更新 HEDGEHOG信号通路 HT-29细胞 

分 类 号：R285[医药卫生—中药学]