关键词:  c-Jun N 端激酶, shRNA;转染, 肝肿瘤, 实验性, 淋巴转移

Abstract:

Abstract:Objective To study the inhibition of  shRNA on  JNK expression  and influence on migration and invasion of mouse hepatocellular carcinoma cell line Hca-F  JNK and  discuss the correlation between expressing level of JNK and lymphatic metastasis of mouse hepatocarcinoma.Methods Three shRNA expression vectors were built and transfected to Hca-F cells stablely.　The inhibition of  shRNA on the expressions of mRNA and protein of JNK  were detected by RT-PCR and Western blotting.The most effective shRNA was selected.Transwell assay was used to detect the inhibition of shRNA on migration and invasion.Results The expression vector of pSilencer-shRNA was built and transfected to Hca-F cells successfully and the most effective shRNA was selected which inhibited JNK expression.The expression of JNK mRNA was markedly decreased after transfection of shRNA  ( P<0.01 ).The expression of JNK protein was markedly inhibited compared with the other groups ( P<0.01 ).The abilities of migration and invasion were decreased after transfection of shRNA ( P<0.05 ).Conclusion The Hca-F cell lines with markedly decreased expression of JNK by stable transfection is obtained by establishing pSilencer-shRNA vector.The inhibition of JNK expression could decrease the abilities of migration and invasion of mouse hepatocellular carcinoma cell lines.JNK maybe play an important role in lymphatic metastasis of hepatocellular carcinoma.

Key words: c-Jun N-terminal kinase, shRNA;transfection, liver neoplasms,experimental, lymphatic metastasis