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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 study the expression patterns of miRNAs in esophageal cancer cell line TE-1 and to provide a reference for the early diagnosis of esophageal cancer. Methods The esophageal cancer cell line TE-1 and normal cell HET-1A were cultured by routine method. The total RNA was extracted and labeled by Cy3 fluorescence. The fluorescence signal was scanned with a scanner. The scanned image was digitized by software Five miRNAs with the most up-regulated and down-regulated genes were screened by RT-PCR and verified by real-time reverse transcription-polymerase chain reaction (RT-PCR). Results A total of 676 differentially expressed miRNAs were screened out, of which 113 were significantly different (all P <0.05), including 76 up-regulated and 37 down-regulated expression. Ten miRNAs with significant difference and high signal intensity Fluorescent quantitative RT-PCR detection, the results consistent with the chip test. Conclusion The miRNAs differentially expressed in esophageal cancer cell line TE-1 and normal cell line HET-1A were screened by gene chip method, which laid the foundation for the further search of new esophageal cancer molecular markers.