结肠癌转移相关潜在微小RNA(miRNA)的分离与鉴定（英文）  

Identification of metastasis-associated microRNAs in colorectal carcinoma cells

作　　者：王燕[1] 张金莉[1] 彭莉萍[1] 张晟春[2] 

机构地区：[1]遵义医学院珠海校区生物化学与细胞分子生物学教研室,广东珠海519041 [2]遵义医学院第五附属医院检验科,广东珠海519041

出　　处：《遵义医学院学报》2015年第5期479-487,共9页Journal of Zunyi Medical University

基　　金：贵州省科技计划项目(NO:黔科合OZ字[2009]2)

摘　　要：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 analyze and compare the effect of human intestinal cancer cell ideology on the expression of microRNA （miRNA） has important implications in understanding the biology of intestinal cancer metasta- sis and in developing new diagnostic and therapeutic agents. Methods Utilizing the combined technology of improved suppression subtractive hybridization （SSH） and backward dot hybridization, we built expression profiles of miRNAs in two different colon cancer cell lines, SW620 and SW480. The miRNAs sequences were compared with the GenBank database using BLAST, we have determined the order and type of miRNA expressed by these cells and used the miRBase to determine the target gene associated with each miRNA. Results Vie identified 24 miRNAs that were differentially expressed between SVi620 and SW480 and found that 4 of those were new, as yet unidentified miRNAs. Some of these miRNAs had previously been shown to be involved in the development of intestinal cancer. Some of the target genes of these miRNAs are involved in gene transcription, cell cycle, signal transduction and cell apoptosis. Our data shows that these miRNAs are involved in the proliferation, invasion and metastasis and so on. These miRNAs have various biological behaviors of colon cancer, and have clearly quantifiable differences in miRNAs expression between human intestinal cancer cell lines SW620 and SW480. These differentially expressed miRNAs may participate in the transfer process of colon cancer. Conclusion The improved subtractive hybridization approach described in this study can be used to effectively discern differential expressions of miRNA in two different specimens. The result of this study offers a new direction for molecular mechanism in the study of metastasis of the colorectal carcinoma.

关 键 词：微小RNA 结直肠癌 转移 抑制性消减杂交 反向斑点杂交技术 

分 类 号：R735.3[医药卫生—肿瘤]