基于Hedgehog信号通路的半枝莲抑制结直肠肿瘤干细胞分化的研究

肿瘤干细胞（CSC）是机体内极小部分能够在病理条件下不对称分化为肿瘤的细胞。我们发现Hedgehog信号通路在CSC自我更新和分化中起有重要作用。研究以Hedgehog信号通路为作用靶点的药物、抑制CSC分化对于防治肿瘤具有重要意义。我们发现半枝莲可以通过下调Hedgehog信号通路活性抑制结直肠CSC自我更新，并抑制CSC异种移植小鼠后分化的肿瘤生长、降低此肿瘤组织中Gli1 mRNA和蛋白水平。本项目采用悬浮培养和3-D培养原代结直肠CSC（pCSC）、分拣自HT-29人结直肠肿瘤细胞的CSC（HT-29CSC）、用CRISPR-Cas9基因组编辑技术稳定敲除Sufu基因的HT-29CSC，并以CSC异种移植NOD/SCID小鼠为模型，研究半枝莲对结直肠CSC分化的影响，从Hedgehog信号通路阐明其抑制CSC分化的机制，为半枝莲防治肿瘤提供科学的实验基础和新的治疗理论。

Cancer stem cells (CSC) in the body are an extremely small portion of cells, which can asymmetrically differentiate into cancer under pathological condition. We found that hedgehog signaling pathway may play a key role in the self-renewal and differentiation of cancer stem cells. Therefore, it is very important to therapeutically target the molecules of hedgehog signaling pathway and prevent cancer stem cells from differentiation. We found that Scutellaria barbata D. Don (SB) can inhibit the self-renewal of colorectal cancer stem cells by down regulation of hedgehog signaling pathway activity, suppress the CSC xenograft tumor growth in mice and reduce the levels of Gli1 mRNA and protein in the tumor. Using suspension culture and 3-D culture of primary human colorectal cancer stem cells (pCSC), cancer stem cells sorted from HT-29 human colorectal adenocarcinoma cell line (HT-29CSC), HT-29CSC with Sufu gene stably knocked out by CRISPR-Cas9 genome editing system and CSC xenograft tumor in NOD/SCID mouse model, we seek to investigate the effect of SB on colorectal cancer stem cells differentiation, and unveil the therapeutic mechanism related to hedgehog signaling pathway, so as to provide scientific support and new insight into the theory for prevention and treatment of cancer with SB.