shRNA抑制小鼠肝癌细胞株JNK的表达及细胞迁移和侵袭

J4 ›› 2009, Vol. 35 ›› Issue (3): 410-414.

shRNA抑制小鼠肝癌细胞株JNK的表达及细胞迁移和侵袭

张宇虹¹| 唐建武²| 王绍清²| 王志强³|孙成荣²|王梅²|王波²

1. 大连医科大学附属第二医院超声科,辽宁大连 116027；2. 大连医科大学病理学教研室|辽宁 |大连 |116044；3. 大连医科大学附属第一医院耳鼻喉科,辽宁大连 116011

收稿日期:2009-01-07 出版日期:2009-05-28 发布日期:2009-08-14
通讯作者: 唐建武 E-mail:jianwutang@ 163.com
作者简介:张宇虹（1966－）| 男| 吉林省榆树市人|教授|在读医学博士|主要从事肝癌转移机制的研究。
基金资助:
国家自然科学基金资助课题（30572098）

Inhibitory effects of shRNA on expression of JNK |in mouse hepatocellular carcinoma cell lines and cell migration and invasion

ZHANG Yu-hong¹,TANG Jian-wu²,WANG Shao-qing²,WANG Zhi-qiang³,SUN Cheng-rong²,WANG Mei²,WANG Bo²

1. Department of Ultrasound,Second Affiliated Hospital, Dalian Medical University,Dalian 116027,China;2. Department of Pathology,Dalian Medical University,Dalian 116044,China；3. Department of Otorhinolaryngology,First Affiliated Hospital, Dalian Medical University,Dalian 116011,China

Received:2009-01-07 Online:2009-05-28 Published:2009-08-14

摘要/Abstract

摘要：

目的：研究shRNA对小鼠肝癌细胞株Hca-F JNK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Hca-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。

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

中图分类号:

R735.7

张宇虹, 唐建武, 王绍清, 王志强, 孙成荣, 王梅, 王波. shRNA抑制小鼠肝癌细胞株JNK的表达及细胞迁移和侵袭[J]. J4, 2009, 35(3): 410-414.

ZHANG Yu-Gong, Tang-Jian-Wu, Wang-Chao-Qing, Wang-Zhi-Jiang, SUN Cheng-Rong, WANG Mei, WANG Bei. Inhibitory effects of shRNA on expression of JNK |in mouse hepatocellular carcinoma cell lines and cell migration and invasion[J]. J4, 2009, 35(3): 410-414.

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