Analysis and clinical application of ubiquitin system as a new target molecule in non-small cell lung cancer.

Pirh 2被克隆为蛋白ARNIP，其具有与雄激素受体连接的环指结构域。据报道，该Pirh 2具有p53的泛素连接酶活性，并导致p53降解。如果Pirh 2的泛素连接酶活性增强抑癌基因产物如p53的分解，则癌症生长将加速。然而，Pirh 2在临床病例中的作用尚不清楚。方法：从非小细胞肺癌组织中提取总RNA，用逆转录酶合成Pirh 2的cDNA，用真实的时间RT-PCR检测Pirh 2的表达。引物设置为外显子8或9以夹持Pirh 2基因的内含子。使用上述引物进行真实的实时RT-PCR。结果：20例腺癌中有13例癌组织中Pirh 2的表达高于正常肺组织2倍以上。高分化腺癌中Pirh 2的表达低于中低分化腺癌，9例鳞癌中有8例癌组织中Pirh 2的表达是正常肺组织的2倍以上，1例肺癌组织中Pirh 2的表达经免疫组化证实。Pirh 2在70%的非小细胞肺癌中呈高表达。据认为，在非小细胞肺癌中，通过Pirh 2的泛素化，抑制性癌基因如p27和p53的分解被增强。细胞肺癌提示Pirh 2可能参与肺癌的发生发展，其与临床特征、预后、抗癌药物敏感试验等的相关性研究有望进一步深入。

Pirh2 was cloned as protein ARNIP which had a RING- finger domain to be connected to an androgen receptor. It is reported that this Pirh2 has ubiquitin ligase activity of p53, and leads p53 to degradation. Cancer growths will be accelerated if resolution of suppressor oncogene products such as p53 is enhanced by ubiquitin ligase activity of Pirh2. However, the roles of Pirh2 in clinical cases were not clear. Therefore we assayed expression of Pirh2 by real time RT-PCR using the non small-cell lung cancer tissues.Methods :The cDNA was composed by reverse transcriptase after getting a total RNA from a tissue. Primer was set to exon eight or nine to sandwich intron of a Pirh2 gene. Real time RT-PCR was performed using the primer mentioned above. In addition, degree of expression of Pirh2 and p27 was analysed by immunostaining.Results :In 13cases out of 20 adenocarcinoma, Pirh2 was more than double in cancer part tissues comparing with normal lung tissue. Well differentiated adenocarcinoma showed less expression of Pirh2 than moderately-poorly differentiated adenocarcinoma.In 8cases out of 9 squamous-cell carcinoma, Pirh2 was more than double in cancer tissues comparing with normal lung tissue.Manifestation of Pirh2 was confirmed in a lung cancer tissue by immunostaining.Discussion :High expression of Pirh2 was present in 70% of non small-cell lung cancer.It is thought that resolution of suppressor oncogene such as p27 and p53 is enhanced by ubiquitination through Pirh2 in non small-cell lung cancer. It was suggested that Pirh2 may contribute to the carcinogenesis or development of lung cancer.It seems that this research can develop concerning a correlation with clinical characteristics, prognosis, anticancer drug sensitivity test and so on.