食管癌细胞株TE-1 microRNA表达谱研究

Profile of microRNA expression in esophageal cancer TE-1 cells

目的研究食管癌细胞株TE-1的微小核糖核酸(miRNAs)表达谱,为食管癌的早期诊断提供参考依据。方法常规方法培养食管癌细胞TE-1和正常细胞HET-1A,提取其总RNA,经Cy3荧光标记后进行片段化、芯片杂交,应用扫描仪扫描芯片荧光信号图像,用软件对扫描图像进行数字化处理和分析,筛选出差异表达miRNA;挑选芯片检测中上调和下调最明显的miRNA各5条,应用实时定量反转录聚合酶链反应(RT-PCR)检测予以验证。结果筛选出差异表达的miRNA 676条,其中113条差异均有统计学意义(均P<0.05),包括76条上调表达和37条下调表达;挑选差异显著且信号强度较高的10条miRNA进行荧光定量RT-PCR检测,其结果与芯片检测一致。结论采用基因芯片方法筛选到食管癌细胞株TE-1与正常细胞株HET-1A差异表达的miRNAs,为进一步寻找新的食管癌分子标志物奠定基础。

Objective To examine the profile of microRNAs （miRNAs）expression in esophageal cancer TE-1 cells and to provide references for early diagnosis of esophageal cancer. Methods The esophageal cancer cells TEA and normal human esophageal cells HET-1A were cultured and their total RNA were isolated. Then, the cDNA was synthesized by reverse transcription and labeled with Cy3 fluorescence as probes,hybridized with microRNA gene chip, and scanned by laser scanner. The acquired image was analyzed for differentially expressed miRNAs. The real time reverse transcription-PCR（RT-PCR） was used for the determination of both five most up- and down-regulated miRNAs. Results A total of 676 miRNAs were screened from the TE-1 line cells ,among which 113 miRNAs were statistically different from those of HET-1A cells （all P 〈 0. 05 ）, including 76 up-regulated and 37 down-regulated miRNAs. The detection results of real time RT-PCR for 10 more differentially expressed miRNAs were consistent with those of gene chip. Conclusion The miRNAs differentially expressed between the esophageal cancer cells lines TE-1 and normal human esophageal cells HET-1A were obtained by miRNAs microarray. The study on these differentially expressed miRNAs could provide a good base for the study on development mechanism of esophageal cancer.