肺腺癌相关基因及其与代谢通路关系的研究

Study of gene expression profile of pulmonary adenoma and its metastasis-related genes and the correlation of the discrepant genes with metabolism pathway

目的利用基因芯片技术研究肺腺癌、转移淋巴结、癌旁正常及胚胎肺组织中的基因表达差异,并利用代谢通路数据库分析这些基因与代谢通路相关性信息。方法分别抽取肺腺癌、转移淋巴结、癌旁正常和胚胎肺组织的总RNA并纯化mRNA,用逆转录的方法,将各mRNA的两种荧光Cy5和Cy3标记的cDNA链做探针,并与含有1152条人类全长基因芯片上进行杂交。用计算机处理和分析。将实验所得基因映射到(Kyoto encyclopedia of genes and genomes,KEGG)通路数据库的基因表达调控通路上,检测同一基因表达调控通路内基因的共表达趋势。结果4例肺腺癌组织和癌旁正常组织比较共表达差异基因25个;3例肺腺癌组织和转移淋巴结组织比较共表达差异基因8个;1例胚胎肺组织和癌旁正常组织比较差异表达基因316个,且与肺腺癌组织和癌旁正常组织比较16个基因有共同表达差异。在β-丙氨酸代谢通路和丁酸代谢通路中的基因与疾病组织中的基因显著相关(P<0.05),而与癌旁组织和淋巴结组织中的基因无关(P>0.05)。结论肺腺癌组织与癌旁正常组织比较、肺腺癌组织和转移淋巴结组织比较均存在共表达差异基因,这些基因可能与肺腺癌的发生、发展有关。代谢通路分析结果提示,在β-丙氨酸代谢通路和丁酸代谢通路中存在着肺腺癌疾病相关的基因,且在同一通路中的基因有共表达趋势。

[Objective] To investigate the differences of gene expression profile among the pulmonary adenoma, metastatic lymph nodes, normal pericancerous and fetal lung tissue by means of cDNA microarray,analyse the correlation of the discrepant genes with KEGG（Kyoto Encyclopedia of Genes and Genomes）database. [Methods] Total RNA was extracted from pulmonary adenoma, metastatic lymph nodes, normal pericancerous and fetal lung tissue in order to get purified mRNA which was retro-transcribed into cDNA. As the probes, the cDNA were hybrided with the whole gene chip of human. By irradiating the obtained genes from the experiment into the KEGG pathway database and analyse the correlation. [Result] In 4 subjects with pulmonary adenoma and normal pericancerous tissue, 25 genes were screened out for common differences in gene expression level. 8 genes in 3 subjects with pulmonary adenoma and metastatic lymph nodes tissue were also identified for common differences. 316 genes were screened out for differences in 1 subject of fetal lung and normal pericancerous tissue. Compared with pulmonary adenoma and normal pericancerous tissue, 16 genes in the subject of pulmonary adenoma and normal pericancerous tissue were screened out for common differences. In β-alanine metabolism pathway and butanoic acid metabolism pathway, the genes in the illness group are notably dependent （P 〈0.05） . [Conclusion] The results demonstrates that the 25 genes and 8 genes are probably related with the occurrence and development of pulmonary cancer; The 316 genes are might related with the developmental circumstances; The 16 genes possibly associated with the initiation of early-stage pulmonary cancer. The results for the experiment data prompt that the genes in β-alanine metabolism pathway and butanoic acid metabolism pathway may jointly take part in the occurrence and development processes for the bronchogenic adenocarcinoma.