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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 study the effects of Scutellaria barbata on the self-renewal of colorectal cancer stem cells (CSCs), and elucidate its mechanism of action from the Hedgehog signaling pathway. Methods: The ethanol extract of Scutellaria barbata was prepared and the colon CSCs were sorted out from HT-29 human colon adenocarcinoma epithelial cells. Aqueous method was used to study the effect of Scutellaria barbata on the activity of colon CSC in vitro. Effect of Zhilian on Colonic CSC Self - renewal. The effect of Scutellaria barbata on CD133 and Lgr5 gene expression in colonic CSC and the expression of Hedgehog signaling gene Ptch1 and Gli1 in colonic CSC were studied by qRT-PCR. Results: ethanol extract of Scutellaria barbata could inhibit the activity of CSC in vitro, inhibit the self-renewal of colon CSC, and downregulate the expression of CD133 and Lgr5 genes in colonic CSC marker in 125μg / ml, 250μg / ml and 500μg / ml Level, reduce Hedgehog signaling pathway genes Ptch1 and Gli1 mRNA transcription in colon CSC. Conclusion: Scutellaria barbata can inhibit the self-renewal of colon CSC by inhibiting the activity of Hedgehog signaling pathway.