鼻咽上皮细胞中过表达PIN1 激活JNK信号通路上调CyclinD1的研究

PIN1 overexpression activates JNK signal pathway and upregulates CyclinD1 in nasopharyngeal epithelium cell

摘要:

目的：研究肽脯氨酰异构酶PIN1（prolylisomerase，PIN1）可能影响的肿瘤信号通路和相关基因，探讨鼻咽癌早期发生发展的机制。方法：运用慢病毒表达系统在鼻咽上皮细胞系 NP69 中建立PIN1 过表达细胞克隆株，real-time PCR、Western blot检测PIN1表达情况；肿瘤信号通路阵列荧光素酶实验筛选PIN1可能影响的细胞信号通路，Western blot加以验证。结果：稳定表达 PIN1 的NP69 细胞可激活一系列肿瘤信号通路其中激活丝裂原活化蛋白激酶MAPK/c-Jun氨基末端激酶JNK（MAPK/JNK） 信号通路，差异具有统计学意义（P<0.01），核转录因子－κB（P=0.036）、癌基因Myc（P=0.048）信号通路变化具有统计学意义（P<0.05）。Western blot结果显示与对照组相比，稳定表达 PIN1 的NP69实验组可明显上调JNK 信号通路重要的下游转录因子c-Jun、c-Jun Ser63、c-JunSer73的表达量，最终上调下游基因细胞周期蛋白CyclinD1（t=-7.295，P=0.002）的表达。结论：稳定表达 PIN1 的NP69 细胞可激活MAPK/JNK信号通路，明显上调JNK 信号通路重要的下游转录因子c-Jun、c-Jun Ser63、c-JunSer73的表达量，最终上调下游基因细胞周期蛋白CyclinD1的表达，PIN1与鼻咽癌早期的发生发展密切相关。

Abstract:

Objective：To investigate the cancer signal pathways which prolylisomerase（PIN1） may regulate. Methods：PIN1 overex－pression NP69 cell line was established by using the leti-viral system and the PIN1 expression was proved by q-PCR and Western blot. Then PIN1 regulated cancer signal pathways were analyzed by using cancer 10-pathway reporter assay and Western blot. Results：PIN1 over expression in nasopharyngeal epithelia cell may activate a serial cancer signal pathways including MAPK/JNK signal pathway，NF-κB signal pathway and Myc signal pathway. Comparing with those control group，Western blot results showed NP69-Lenti-PIN1 group had higher quantity expressions in c-Jun，c-Jun Ser63，c-JunSer73 and CyclinD1（t=-7.295，P=0.002）. Conclusion：The over-expression of PIN1 up-regulates CyclinD1 expression，activates the MAPK/JNK pathway，and increases phosphorylation of c-Jun at Ser63 and Ser73. Therefore PIN1 may contribute to the tumorigenesis of nasopharyngeal carcinoma.