细胞色素氧化酶10在非梗阻性无精子症患者睾丸组织中的表达

Expression of COX10 in Human Non-Obstructive Azoospermia Testes

目的:评价细胞色素氧化酶10(COX10)基因在非梗阻性人无精子症及正常睾丸组织中的表达及意义。方法:应用包含有人COX10及RBM、EIF1AY等基因的微矩阵芯片对人正常睾丸及无精子症睾丸组织中差异表达基因谱进行了研究:通过RT-PCR方法获得两种组织mRNA,再分别用Cy5-dUTP及Cy3-dUTP标记制备cDNA探针。两种探针混合后与包含有人4096条cDNA序列的cDNA微矩阵芯片杂交,经扫描、计算机处理分析比较杂交结果;随后利用原位杂交技术对COX10mRNA在10例正常生育及39例非梗阻性无精子症睾丸组织中的表达进行了研究。结果:获得128个可能与无精子症相关的差异表达基因,其中56个基因表达上调,72个基因表达下调,其中COX10下调明显。与cDNA微矩阵杂交结果相同,原位杂交证实COX10mRNA在正常睾丸组织生精细胞中表达强于非梗阻性无精子症睾丸组织。结论:COX10可能在无精子症的发生与进展过程中起一定的作用;cDNA微矩阵技术可以应用于筛选非梗阻性无精子症相关基因的研究中。

Objective： To evaluate the expression of COXIO mRNA in the testes of non-obstructive azoospermia patients and normal men. Methods： A cDNA microarray containing COX10 and some other genes as RBM and EIF1AY was used to identify the differential gene expression profiles in the normal and azoospermic testes. The cDNA probes were prepared by labeling mRNA from azoospermic and normal testis tissues with CyS-dUTP and Cy3-dUTP respectively through reverse transcription. The mixed cDNA probes were then hybridized with cDNA microarray. Later the fluorescent signals were scanned and the values of CyS-dUTP and Cy3- dUTP on each spot were calculated and analyzed. After that an ISH was employed to detect the expression of COX10 mRNA in 10 fertile and 39 non-obstructive azoospermic testes, and the expression levels were compared to evaluate the significance. Results ： We obtained 128 differentially expressed genes that might be related with azoospermia, among which 56 were up-regulated and 72 down-regu- lated, with the expression of COXIO significantly decreased. In situ hybridization confirmed that the mRNA expression of COX10 was stronger in the spermatogenic cells of the normal fertile than the azoospermic testes. Conclusion ： COX10 may play a certain role in the development and progression of azoospermia. The technique of cDNA microarray can be applied to further studies of screening nonobstructive azoospermia associated genes.