基于Hedgehog信号通路的野黄芩苷抑制HT-29细胞源性结肠肿瘤干细胞体外自我更新的研究  

Inhibition of the self-renewal in vitro of cancer stem-like cells in HT-29 cells by scutellarin via down-regulation of hedgehog signaling pathway activity

作　　者：熊思会 万莎 谭溧 雷楠 卢小路 沈小钢[2] 周丽娟[3] 徐海波[1] 

机构地区：[1]成都中医药大学药学院药理系,成都611137 [2]四川省人民医院胃肠外科,成都610072 [3]四川省中医药科学院,成都610014

出　　处：《中药药理与临床》2017年第5期13-16,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(编号:81573813;81173598);教育部留学回国人员科研启动基金(编号:2013693)

摘　　要：目的:研究半枝莲中主要成分野黄芩苷对结肠肿瘤干细胞(CSC)自我更新的影响,并从Hedgehog信号通路阐明其作用机制。方法:从HT-29人结肠腺癌上皮细胞株中分拣出结肠肿瘤干细胞(HT-29CSC),并对其进行无血清悬浮培养。用CCK-8试验研究野黄芩苷对HT-29CSC活性的影响;用干细胞球形成试验研究野黄芩苷对HT-29CSC体外自我更新的影响;用qRT-PCR研究野黄芩苷对结肠肿瘤干细胞标记物基因CD133和增殖基因ki-67表达的影响,和对结肠肿瘤干细胞中Hedgehog信号通路基因Ptch1和Gli1表达的影响。结果:终浓度为40μg/ml、80μg/ml和160μg/ml的野黄芩苷可以浓度依赖性地抑制HT-29CSC体外活性,抑制HT-29CSC体外自我更新,下调结肠肿瘤干细胞标记物基因CD133和增殖基因ki-67 mRNA水平,抑制结肠肿瘤干细胞中Hedgehog信号通路基因Ptch1和Gli1 mRNA转录。结论:野黄芩苷可以抑制结肠肿瘤干细胞活性和自我更新,其机制在于抑制Hedgehog信号通路的活性。Objective： To investigate the effect of scutellarinon on the self-renewal of colonic cancer stem cells（CSC） and its underlying mechanism through hedgehog signaling pathway. Methods： Colonic cancer stem cells were fractionated from human colonic adenocarcinoma epithelial cells HT-29,named HT-29 CSC. The effect of scutellarin on the cell viability of HT-29 CSC was determined with CCK-8 assay. The function of scutellarinon HT-29 CSC self-renewal was assessed with spheroid formation assay. The effect of scutellarin on expressions of colonic CSC marker gene CD133 and proliferation gene Ki-67,and expressions of Ptch1 and Gli1 involved in hedgehog signaling pathway of colonic CSC were measured by quantitative real-time RT-PCR. Results： Scutellarin at final concentrations of 40μg/ml,80 μg/ml and 160 μg/ml dosedependently inhibited HT-29 CSC viability and its self-renewal,down regulated mRNA levels of CD133 and Ki-67,and decreased mRNA transcriptions of Ptch1 and Gli1 involved in hedgehog signaling pathway of colonic CSC. Conclusions： Scutellarin suppresses colonic CSC selfrenewal by reducing the activity of hedgehog signaling pathway.

关 键 词：野黄芩苷 肿瘤干细胞 自我更新 HEDGEHOG信号通路 

分 类 号：R285[医药卫生—中药学]