中文摘要：

目的：研究shRNA对小鼠肝癌细胞株Hca—FJNK基因表达的抑制作用及对肝癌细胞迁移和侵袭能力的影响，阐明JNK表达水平与小鼠肝癌细胞淋巴转移潜能的关系。方法：构建3条shRNA表达载体，以脂质体法稳定转染Hea—F细胞；RT-PCR和Western blotting检测shRNA对JNK基因和蛋白表达的抑制作用，筛选出RNA干扰效果最好的shRNA；Transwell实验检测Hca-F细胞穿膜细胞数观察shRNA对Hca—F细胞迁移和侵袭能力的抑制作用。结果：成功构建pSilencer-shRNA表达载体并稳定转染Hca—F细胞，筛选出对JNK表达抑制效果最好的shRNA；其转染后JNK mRNA表达水平与其他组比较明显下调（P〈0．01）；JNK蛋白质表达水平与其他组比较明显减少（P〈0．01）；转染shRNA后Hca—F细胞穿膜细胞数均显著下降（P〈0．05）。结论：通过建立psilencer-shRNA表达载体并稳定转染Hca—F细胞株，获得JNK表达稳定下调的Hca—F细胞株，抑制JNK表达水平可降低小鼠肝癌细胞的迁移和侵袭能力，JNK可能是决定肝癌淋巴转移潜能的重要基因之一。

英文摘要：

Objective To study the inhibition of shRNA on JNK expression and influence on migration and invasion of mouse hepatocellular carcinoma cell line Hea-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 transfeetion 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.