从细胞基因表达谱分析肾精不足与恶性肿瘤的相关性

Analysis of the Correlation Between Kidney Essence Deficiency and Malignant Tumors by Cell Gene Expression Profile

目的:通过检测肾精不足证和Lewis肺癌肺实质细胞基因表达,比较二者细胞相应基因表达,分析肾精不足是否参与肺癌细胞基因变异。方法:分离正常小鼠、肾精不足证小鼠和Lewis肺癌小鼠肺上皮细胞,提取总RNA,扩增染色后,与鼠全基因组芯片杂交,排除肾精不足证小鼠和Lewis肺癌小鼠二者与正常小鼠基因中相似成分,利用生物基因筛选技术对二者剩余表达基因进行筛选,获得同源表达基因后进行GO分析和基于KEGG数据库的基因通路分析以及qRT-PCR验证。结果:GO分析表明:二者细胞组分表达基因相似度为26%,其中胞内非膜结合细胞器胞膜和胞外区比例最高;细胞分子功能表达基因相似度为33%,其中激酶活性、转移酶活性和信号转导调节比例最高;生物过程的表达基因相似度为39%,其中细胞周期、生物调节、细胞分化比例最高。KEGG通路分析显示:mTOR信号通路、PI3K/Akt、wnt通路有明显相似性,共涉及12个相似基因表达。qRT-PCR结果与芯片结果比较,差异无统计学意义(P>0.05)。结论:肾精不足证小鼠与肺癌小鼠上皮细胞基因表达谱存在相似性,且主要体现在细胞生命过程中,如细胞分化、周期调节以及细胞功能等。这也与中医肾有推动人体生长发育的功能有关。

Objective:To analyze whether kidney essence deficiency is involved in gene mutation in lung cancer cells by detecting the gene expression in lung parenchymal cells of kidney essence deficiency syndrome and Lewis lung cancer and comparing the corresponding gene expression of the two cells.Methods:Total RNA was extracted from lung epithelial cells of normal mice,kidney essence deficiency mice and Lewis lung cancer mice.After amplification and staining,the cells were hybridized with whole genome microarray of mice,excluding the similar components in the genes of kidney essence deficiency mice and Lewis lung cancer mice and normal mice.The remaining expressed genes of the two mice were screened by biotechnology.The homologous expressed genes were screened,then GO analysis,gene pathway analysis based on KEGG database and qRT-PCR validation were carried out.Results:GO analysis showed that the similarity of gene expression between the two cell components was 26%,of which the highest proportion was found in the membranes and extracellular regions of intracellular non-membrane-bound organelles,33%in the functional expression genes of cells,and 39%in kinase activity,transferase activity and signal transduction regulation.The proportion of cell cycle,biological regulation and cell differentiation was the highest.KEGG pathway analysis showed that the mTOR signaling pathway,PI3 K/Akt,Wnt pathway had obvious similarities,involving 12 similar gene expression.There was no significant difference between qRT-PCR and microarray results(P>0.05).Conclusion:The gene expression profiles of epithelial cells of mice with kidney essence deficiency syndrome are similar to those of lung cancer mice,and are mainly reflected in the process of cell life,such as cell differentiation,cell cycle regulation and cell function.This is also related to the function of TCM kidney in promoting human growth and development.