结直肠癌中微小RNA调控网络识别

发布时间：2019-06-27 20:36

【摘要】：MicroRNA是一类约22bp的非编码单链RNA分子其可以在转路后水平调节基因的表达,这些小分子可以在真核基因表达调控中发挥着非常重要的作用。并且,miRNA的表达模式具有分化的位相性和时序性。但是miRNA在结直肠癌中的调控网络依然存在很多不明了的环节,因此,在本文中我们选取了两个在结直肠癌中具有重要作用的miRNA,并对其表达调控机制进行了研究。首先我们利用miRNA测序和Northern Blot的方法检测到miR-145和miR-143在结直肠癌中表达显著下调,相反,如果在结直肠癌细胞中过表达miR-145和miR-143则明显抑制肿瘤细胞的增殖,细胞浸润及成瘤能力。进一步研究中,我们通过自行研发的miRNA靶标预测软件KeyTar发现miR-143的潜在靶标是受体酪氨酸蛋白激酶ErbB3口GTP酶K-ras,而mi R-145的潜在靶标是N-ras。随后我们通过萤光素酶报告实验进一步验证了这个结论,同时,Western Blot结果表明miR-143的高表达可以抑制ErbB3和K-ras的表达,miR-145的高表达可以抑制N-ras的表达,这些结论也表明了miR-143和miR-145通过对靶基因的抑制作用从而对结直肠癌的发生和发展起到重要作用。
[Abstract]:MicroRNA is a class of non-coding single-stranded RNA molecules about 22bp, which can regulate gene expression at the level after transposition. These small molecules can play a very important role in the regulation of eukaryotic gene expression. Moreover, the expression pattern of miRNA has the phase and timing of differentiation. However, there are still many unclear links in the regulatory network of miRNA in colorectal cancer. Therefore, in this paper, we selected two miRNA, which play an important role in colorectal cancer and studied its expression and regulation mechanism. Firstly, we detected that the expression of miR-145 and miR-143 in colorectal cancer was significantly down-regulated by miRNA sequencing and Northern Blot. On the contrary, if miR-145 and miR-143 were overexpressed in colorectal cancer cells, the proliferation, cell invasion and tumorigenesis of colorectal cancer cells were significantly inhibited. In further study, we found that the potential target of miR-143 was GTP enzyme K 眉 rasase of receptor tyrosine protein kinase ErbB3, and the potential target of miR- 145 was N 眉 ras. through the miRNA target prediction software developed by ourselves, we found that the potential target of miR-143 was GTP enzyme K 眉 rasase in ErbB3 port of receptor protein kinase. Then we further verified this conclusion by luciferase reporting experiment. At the same time, Western Blot results showed that the high expression of miR-143 could inhibit the expression of ErbB3 and K-ras, and the high expression of miR-145 could inhibit the expression of N-ras. These conclusions also indicated that miR-143 and miR-145 played an important role in the occurrence and development of colorectal cancer by inhibiting the target gene.
【学位授予单位】：华东师范大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R735.34

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