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肿瘤微环境中肝星状细胞能够影响肝癌细胞的生物学行为.在肝脏发育过程中,肝星状细胞通过表皮形态发生素(epimorphin,EPM,syntaxin2)与肝干细胞接触而促进肝干细胞的功能性分化.EPM在溃疡性结肠炎、间质性肺炎以及结肠癌中也发挥了重要的作用.发现肝细胞癌(HCC)细胞能够促使星状细胞EPM表达上调.为了研究肝星状细胞的EPM对于肝癌可能的作用,构建了针对EPM基因表达的shRNA干扰载体,并将质粒转染肝星状细胞,获得了两株携带该干扰片段的细胞系.RT-PCR与Westernblot检测结果表明,转基因干扰星状细胞系EPMmRNA和蛋白质表达量明显降低.之后,使用转基因细胞系的条件培养基对于肿瘤细胞进行侵袭能力的检测,并对星状细胞与肝癌细胞三维共培养,证明EPM干涉后肝星状细胞促进肿瘤细胞迁移的能力降低.结果表明通过RNAi可稳定干扰人肝星状细胞EPM基因的表达,并且EPM能够促进肝癌细胞的转移.
Hepatic stellate cells can affect the biological behavior of hepatocellular carcinoma cells in tumor microenvironment. During hepatic development, hepatic stellate cells promote hepatic stem cell function through contact with hepatic stem cells through epimorphin (EPM, syntaxin2) Differentiation.EPM also plays an important role in ulcerative colitis, interstitial pneumonia and colon cancer.Hepatocellular carcinoma (HCC) cells were found to promote the upregulation of stellate cell EPM.In order to study the effect of EPM on hepatic stellate cells The possible role of RNA interference in the development of hepatocellular carcinoma and the construction of a shRNA interference vector targeting the expression of EPM gene and transfecting the plasmid into hepatic stellate cells resulted in two cell lines carrying the interference fragment.The results of RT-PCR and Western blot showed that transgene interfering star The expression level of EPMmRNA and protein in the cell line decreased significantly.After that, the invasion ability of the tumor cells was detected by using the conditioned medium of the transgenic cell line, and the three-dimensional co-culture of the stellate cells and the liver cancer cells was performed to prove that the hepatic stellate cells The ability to promote the migration of tumor cells is reduced.The results show that RNAi can stably interfere with the expression of EPM gene in human hepatic stellate cells and EPM can Transfer into liver cancer cells.