茎-环反转录实时PCR技术分析miR-93和miR-342在乳腺癌细胞系中的表达  

Expression analysis of miR-93 and miR-342 in breast cancer cell lines by stem-loop reverse transcription real-time PCR technique

作　　者：杨璐全[1] 杨曼曼[1] 刘晓燕[1] 龙燕[1] 魏春莉[1] 杨维婵[1] 傅俊江[1] 

机构地区：[1]泸州医学院医学基础研究中心表观遗传与肿瘤重点实验室,四川泸州646000

出　　处：《泸州医学院学报》2013年第4期315-319,共5页Journal of Luzhou Medical College

基　　金：国家自然科学基金(编号:81172049);四川省"肿瘤表观遗传学"高校科技创新团队(编号:13TD0032)

摘　　要：目的:应用高灵敏度的特异性茎-环反转录实时PCR技术检测分析小的非编码分子miR-342和miR-93在乳腺癌细胞中的表达。方法:用miR特异性试剂盒(miRNeasy Mini Kit)提取了总RNA和miRNA,进行特异性茎-环反转录和实时PCR(stem-loop reverse transcription real-time PCR,stem-loop RT-qPCR),检测miR-342和miR-93在5种乳腺癌细胞中的表达,并进行比较分析。结果:用高灵敏度的特异性茎-环反转录实时PCR技术检测发现miR-93在T47D细胞系中呈非常高的表达,BT-549和MCF7细胞系中呈相对高表达,在MDA-MB-231和MDA-MB-435细胞系低表达;而miR-342在MCF7中呈非常高的表达,MDA-MB-231,BT-549和T47D中呈相对低表达,在MDA-MB-435中表达特别低。结论:miR-93和miR-342的表达在某种程度上与乳腺癌细胞高侵袭性呈一定负相关,表明miR-93和miR-342可能是乳腺癌的抑癌基因并可能具有抑制其转移的作用,为深入分析miR-93和miR-342抑制乳腺癌转移的作用机制奠定了基础。Objective： To analyze the expression of small non-coding microRNA （miRNA） molecules miR-93 and miR-342 in breast cancer cells by stem-loop reverse transcription real-time PCR technique. Methods：Total RNAs, including miRNAs, were extracted using miRNeasy Mini Kit. The expression of miR-93 and miR-342 were analyzed using stem-loop primers for reverse transcription followed by real-time PCR in five breast cancer cell lines. Results： By stem-loop RT-qPCR technique, the expression level of miR-93 was very high in breast cancer cell line T47D, relatively high in BT-549 and MCF7, but low in MDA-MB-231 and MDA-MB-435;whereas the expression level of miR-342 was very high in MCF7, low in MDA-MB-231, BT-549 and T47D, and very low in MDA-MB-435. Conclusion： Our results provide evidence that the expression levels of miR-93 and miR-342 are to a certain degree negatively correlated with high aggressiveness of cancer cells, indicating that miR-93 and miR-342, as antioncogenes, may play important roles in suppressing tumor metastasis in breast cancer. These findings might lay a foundation for further studying the mechanism of miR-93 and miR-342 in suppressing breast cancer metastasis.

关 键 词：茎-环反转录 实时PCR miR-93 miR-342 乳腺癌细胞系 表达 

分 类 号：R394.33[医药卫生—医学遗传学]