老年人睾丸组织退化的基因表达谱研究

Studies on the testis regression related gene profile in aged male

目的 :研究老年人睾丸组织基因表达谱变化 ,为研究老年人睾丸功能衰退机制奠定基础。方法 :知情同意下获取正常青年人和老年人睾丸组织标本各 3例 ,采用QIAGENRNA提取试剂盒提取总RNA ,分别等量混合后制备探针。利用ClonTech公司生产的包含 80 0 0个已知基因的人类cDNA基因芯片 [AtlasPlasticHuman 8KMi croarray (Cat.# 790 5 1) ]筛选差异表达基因。按基因功能分类方法对老年睾丸组织中差异表达的基因进行归类分析。用RT -PCR、T -A克隆测序等方法选择目的基因 ,验证基因芯片结果。结果 :筛选到差异表达基因 117个(1.4 6 % ) ,83个基因在老年睾丸组织中表达下调 ,34个基因表达上调。在表达下调的基因中 ,代谢相关基因 16个(19.3% ) ,与基因和蛋白表达相关的基因 18个 (2 1.7% ) ,信号通路相关基因 16个 (19.3% ) ,细胞分化相关基因19个 (2 2 .9% ) ,细胞结构和运动相关基因 6个 (7.2 % ) ,细胞或内环境稳态相关基因 4个 (4 .8% ) ,未知功能基因 4个 (4 .8% )。在表达上调的基因中 ,代谢相关基因 3个 (8.8% ) ,与基因和蛋白表达相关的基因 10个 (2 9.4 % ) ,信号通路相关基因 2个 (5 .9% ) ,细胞分化相关基因 11个 (32 .4 % ) ,细胞结构和运动相关基因 8个 (2 3.5 % )。RT -PCR及T

Objective: To understand the mechanism of testis regression in aging male Methods: Human testis tissues were obtained during related operation with informed consent (normal young male 3 cases and aged male 3 cases). Total RNA was isolated by QIAGEN RNAeasy kit. Differentiations of gene expression were studied by Clon Tech cDNA microarray methods and the differential expression gene in aged male were classified by Venter's classify system and the candidate genes were investigated by RT-PCR analysis. Results: In the results of cDNA microarray we found 117(1.46%) gene differentiations in aged male at least more than 1.0 fold. Among them, the 83 genes were down regulated and the 34 genes up regulated. The down expressed genes related to metabolism were 16(19.3%),gene or protein expression 18(21.7%),cell signaling or cell communication 16(19.3%),cell division 19(22.9%),cell structure or motility 6(7.2%)and unknown function 4 genes (4.8%). The up expressed genes related to cell division were 11(32.4%), gene or protein expression 10(29.4%)and metabolism 3 (8.8%). It is interesting to find that respiratory chain related gene cox7a2 was up regulated and atp50 down regulated significantly which as further confirmed by RT-PCR analysis with sequence analysis in the products of the RT-PCR by T-A cloning. Conclusion: The gene expression profile in aged male testis was changed significantly as compared with that in normal young controls; testis regression in aging male may relate multi gene differentiations, especially the differentiations of respiratory chain related gene cox7a2 , atp50 , which may be an important candidate gene in the study of the mechanism of testisregression in aging male.