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目的:分析piRNA在胃癌组织与正常胃黏膜组织中表达的差异性。方法:通过新一代高通量测序技术Solexa对胃癌组织和正常胃黏膜组织进行小分子RNA(s RNA)深度测序,并通过生物信息学分析胃癌组织与正常胃黏膜组织中piRNA的表达差异。根据分析结果选取4种piRNA(登录号分别为DQ570956、DQ575659、DQ594126和DQ597128),通过茎环反转录实时定量PCR(简称茎环RT-q PCR)技术验证其存在及其在胃癌组织和正常胃黏膜组织中的表达差异。结果:Solexa深度测序结果显示,正常胃黏膜与胃癌组织中测得的piRNA种类在各标本所测得所有s RNA种类中所占的比例差异无统计学意义(Z=0.835,P=0.678);正常胃黏膜组织中测得的piRNA数量在各标本所测得s RNA总量中所占的比例高于胃癌组织,差异有统计学意义(Z=2.167,P=0.042);根据分析结果选取4种piRNA,经茎环RT-q PCR法检测显示,这4种piRNA在胃癌组织中的表达量显著低于正常胃黏膜组织,差异有统计学意义(P<0.05)。结论:胃癌组织存在多种差异表达的piRNA,且在表达量上与正常胃黏膜组织存在显著性差异,本研究选取的4种piRNA可能参与胃癌的发生发展进程。
Objective: To analyze the difference of piRNA expression in gastric cancer and normal gastric mucosa. Methods: Solexa was used to perform deep sequencing of small RNA (s RNA) in gastric cancer tissues and normal gastric mucosa tissues. The expression of piRNA in gastric cancer tissues and normal gastric mucosa tissues was analyzed by bioinformatics. Four kinds of piRNAs (accession numbers DQ570956, DQ575659, DQ594126 and DQ597128) were selected according to the results of the analysis, and their existence and their roles in gastric cancer tissues and normal tissues were verified by real-time quantitative PCR of stem-loop reverse transcription Differences in gastric mucosal expression. Results: The results of Solexa sequencing showed that there was no significant difference in the percentage of all RNA species detected in each specimen between normal gastric mucosa and gastric cancer tissues (Z = 0.835, P = 0.678). The number of piRNAs measured in normal gastric mucosa tissues was higher than that in gastric cancer tissues (Z = 2.167, P = 0.042). According to the results, 4 The results of RT-qPCR showed that the expression of these four kinds of piRNAs in gastric cancer tissues was significantly lower than that in normal gastric mucosa (P <0.05). CONCLUSIONS: There are many differentially expressed piRNAs in gastric cancer tissues, and there is a significant difference in the expression level between them and normal gastric mucosa tissues. The four kinds of piRNAs selected in this study may be involved in the development and progression of gastric cancer.