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目的:研究弥漫大B淋巴瘤(Diffuse Large B-Cell Lymphoma,DLBCL)12号染色体基因表达情况。方法:收取临床DLBCL病人淋巴结标本液氮速冻,快速冷冻切片,采用激光显微切割技术分离单纯淋巴瘤细胞,提取淋巴瘤细胞中的mRNA与表达谱芯片杂交,通过信号扫描、处理后获得表达基因杂交信号强度。每基因设11-20对探针。杂交信号与错配探针对比,扣除背景值后,使用Wilcoxon符号秩和检验选取与错配杂交信号有显著差异的基因作为分析结果(P=0.05)。随机选取两个检测到的基因,使用PCR方法检验基因芯片结果的可靠性。结果:成功地从快速冷冻保存的DLBCL标本中提取了RNA。使用表达谱芯片进行研究,发现了共164条12号染色体编码的基因在淋巴瘤细胞中表达。并根据胞内定位,基因功能和基因所属的代谢通路三种分类方法对所得基因进行分类分析。基因表达密度分析显示12号染色体上的基因表达情况与编码基因分布情况比较一致。结论:使用表达谱芯片研究了12号染色体上的基因表达情况,为研究DLBCL提供了依据。
Objective: To study the gene expression on chromosome 12 of Diffuse Large B-Cell Lymphoma (DLBCL). Methods: Lymphoid samples of DLBCL patients were collected and frozen in liquid nitrogen. The lymphoma cells were isolated by laser microdissection. The mRNA of lymphoma cells was extracted and hybridized with the expression profiling chip. After signal processing, the expressed genes were obtained Hybridization signal strength. 11-20 pairs of probes per gene. After subtracting the background value from the hybridization signal and the mismatch probe, the Wilcoxon signed-rank test was used to select the gene which has significant difference with the mismatch hybridization signal (P = 0.05). Two randomly selected genes were tested and PCR assays were used to test the reliability of the gene chip results. Results: RNA was successfully extracted from the cryopreserved DLBCL specimens. Using expression profiling chips, a total of 164 genes encoded by chromosome 12 were found to be expressed in lymphoma cells. According to the three kinds of classification methods of intracellular localization, gene function and gene metabolic pathway, the obtained genes were classified and analyzed. Gene expression density analysis showed that the gene expression on chromosome 12 was more consistent with the distribution of coding genes. Conclusion: The gene expression profile on chromosome 12 was studied by using the expression profile microarray, which provided a basis for the study of DLBCL.