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

Pirh2克隆为ARNIP蛋白，具有一个连接雄激素受体的RING- finger结构域。据报道，该Pirh2具有p53泛素连接酶活性，并导致p53降解。如果pih2的泛素连接酶活性增强了抑制癌基因产物（如p53）的分解，那么癌症的生长将会加速。然而，Pirh2在临床病例中的作用尚不清楚。因此，我们采用实时RT-PCR技术检测了非小细胞肺癌组织中Pirh2的表达。方法：从组织中提取总RNA后，用逆转录酶合成cDNA。引物设置在pih2基因的第8或第9外显子上，夹在内含子上。使用上述引物进行实时RT-PCR。免疫染色法分析Pirh2和p27的表达程度。结果：20例腺癌中，13例癌部组织中Pirh2含量较正常肺组织高2倍以上。高分化腺癌中pih2的表达低于中低分化腺癌。在9例鳞状细胞癌中，有8例癌组织中Pirh2含量是正常肺组织的2倍以上。免疫染色证实了pih2在肺癌组织中的表达。讨论：Pirh2在70%的非小细胞肺癌中高表达。据认为，在非小细胞肺癌中，p27和p53等抑制癌基因的降解通过pih2泛素化而增强。提示Pirh2可能参与肺癌的发生和发展。该研究与临床特点、预后、抗肿瘤药物敏感性试验等方面有一定的联系。

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.