论文部分内容阅读
微小RNA(microRNA)在肿瘤的发生发展中发挥重要作用。有研究表明,在结肠癌患者肿瘤组织中,miR-31表达水平增高。然而,定量PCR只能检测所在组织miR-31的整体表达水平,而无法观察miR-31在特定组织与特定细胞中的表达分布。目前,尚未见关于miR-31在结肠癌中原位表达的报道。本文从研究miR-31在结肠癌中的原位表达入手,进一步探究miR-31在结肠癌细胞中的功能及作用机制。原位杂交实验结果显示,miR-31在结肠癌肿瘤细胞中的原位表达明显升高;体外过表达或敲减miR-31证实,其可以促进结肠癌细胞增殖和集落形成;荧光定量PCR与Western印迹和双荧光素酶报告基因实验证实,在结肠癌细胞中,NF-κB通路的抑制因子丝氨酸/苏氨酸激酶40(STK40)是miR-31下游靶基因,miR-31靶向作用于STK40而激活NF-κB通路;反之,抑制NF-κB通路,miR-31的促增殖能力明显下降。上述结果提示,miR-31可能通过激活NF-κB信号通路而促进结肠癌的细胞增殖。
MicroRNA plays an important role in tumorigenesis. Studies have shown that in patients with colon cancer, miR-31 expression levels increased. However, quantitative PCR only detects the overall expression level of miR-31 in the tissues, but can not observe the expression distribution of miR-31 in specific tissues and specific cells. At present, no report on the expression of miR-31 in situ in colon cancer has been reported yet. In this paper, the study of miR-31 in situ expression in colon cancer to start, further explore the function of miR-31 in colon cancer cells and mechanism of action. In situ hybridization results showed that the expression of miR-31 in colorectal cancer cells significantly increased in situ; miR-31 overexpression or knockdown in vitro confirmed that it can promote colon cancer cell proliferation and colony formation; Fluorescent quantitative PCR and Western blotting and dual-luciferase reporter assay confirmed that miR-31 targets miR-31 target in human colon cancer cells, which is the downstream target of miR-31, a serine / threonine kinase 40 inhibitor of NF-κB pathway STK40 activation of NF-κB pathway; the contrary, inhibition of NF-κB pathway, miR-31 promote proliferation decreased significantly. The above results suggest that miR-31 may promote cell proliferation of colon cancer by activating NF-κB signaling pathway.