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目的构建在两种结肠癌细胞株SW480和SW620中微小RNA的差异表达文库,分离与结肠癌转移具有潜在相关性的微小RNA。方法以具有不同转移潜能的结肠癌细胞株SW480和SW620为材料,应用改良抑制性消减杂交联合反向斑点杂交技术,分离在两种细胞中差异表达的微小RNA,利用生物软件BLAST及MIREAP将分离到的微小RNA序列进行分析比对。结果鉴定出在SW480和SW620之间表达有明显差异的24种微小RNA,其中4种为新发现的微小RNA,并提交至Gen Bank数据库。这些微小RNA中经文献查询已知部分参与了结肠癌的发生发展过程,部分在基因转录、细胞周期循环、信号传导和细胞凋亡过程具有一定的功能。结论经过改良的抑制消减杂交技术可以高效地分离不同标本间差异表达的微小RNA;本研究得出的结果可为结肠癌细胞转移的分子生物学机制研究提供新的方向。
Objective To construct a differential expression library of microRNAs in two colon cancer cell lines, SW480 and SW620, to isolate microRNAs potentially associated with colon cancer metastasis. Methods The colon cancer cell lines SW480 and SW620 with different metastatic potentials were used as materials. The differentially expressed microRNAs isolated from the two cells were isolated by using improved suppression subtractive hybridization and reverse dot blot hybridization. BLAST and MIREAP were used to separate the microRNAs. The microRNA sequence is analyzed and aligned. Results Twenty-four microRNAs were identified that showed significant differences between SW480 and SW620, four of which were newly discovered microRNAs and submitted to the Gen Bank database. Some of these microRNAs are known to have been involved in the development and progression of colon cancer and some have some functions in gene transcription, cell cycle, signal transduction and apoptosis. Conclusion The modified subtractive hybridization technique can efficiently separate microRNAs differentially expressed in different samples. The results obtained in this study may provide a new direction for the molecular mechanism of colon cancer cell metastasis.