液相杂交检测B细胞淋巴瘤细胞系miR-28的表达

Expression of miR-28 in B Cell Lymphoma Cell Lines Detected by Solution Hybridization

microRNA(miRNA)是一组在进化上高度保守、非编码蛋白、参与转录后调节的小分子RNA(19-25核苷酸)。为了进一步明确miRNA的加工机制及功能和定量研究miRNA的表达水平,应用液相杂交和Northernblot两种方法检测miR-28在B细胞淋巴瘤细胞系中的表达并进行了比较。结果表明:液相杂交和Northernblot检测miR-28均显出阳性结果,但液相杂交所用的细胞总RNA量(5μg)明显少于Northernblot(30μg),液相杂交的条带信号也较Northernblot的强。结论:液相杂交是一种较Northernblot更为快速、简便和敏感的检测miR-28的方法。

microRNA （miRNA） is evolutionarily conserved, non-coding small RNA （19 -25 nt） involved in posttranscriptional gene regulation. To further understand the biogenesis and functions of miRNA, and to quantify miRNA expression levels, the expression of miR-28 in B lymphoma cell lines was detected by solution hybridization and Northern blot, and their detected results were compared. The results indicated that detection of miR-28 by solution hybridization and Northern blot showed positive, but total RNA amount used for solution hybridization detection was significantly lower than that for Northern blot detection （5 μg vs 30 μg）, signal of solution hybridization was stronger than that of Northern blot. It is concluded that the solution hybridization for detection of miR-28 in B cell lymphoma cell lines is a faster and more sensitive method as compared with standard Northern blot technique.