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含有乙型肝炎病毒X蛋白的肝癌细胞具有条件性增强的侵袭潜能。

Hepatocellular carcinoma cells containing hepatitis B virus X protein have enhanced invasive potential conditionally.

作者信息

Ou D-P, Tao Y-M, Chang Z-G, Tang F-Q, Yang L-Y

机构信息

Liver Cancer Laboratory, Department of Surgery, Xiangya Hospital, Central South University, Changsha 410008, Hunan, China.

出版信息

Dig Liver Dis. 2006 Apr;38(4):262-7. doi: 10.1016/j.dld.2005.10.027. Epub 2006 Feb 3.

Abstract

AIMS

To establish a sustaining hepatitis B virus X protein expressed Chang liver cell line and to explore their biological behaviours of invasive potential induced by hepatitis B virus X protein.

METHODS

Polymerase chain reaction was used to amplify the HBx gene from the whole hepatitis B virus genome. The gene was then subcloned into the eukaryotic expression vector pcDNA3.1 to construct the pcDNA3.1-HBx plasmid. Gene transfection mediated by Lipofectamine was used to introduce the plasmid into the human liver cell line Chang, and stable expression of the HBx gene was detected.

RESULTS

HBx gene was cloned from the transfected Chang liver cells by reverse transcription-polymerase chain reaction, and confirmed by electrophoresis. The stably transfected Chang cells expressing HBx with malignant characteristics were verified and compared with control cells in terms of their growth curves, clonogenicity, wound healing abilities, migration and metastasis.

CONCLUSION

The stabilising human liver cell lines Chang liver containing HBx gene expression have been established successfully. The invasive potential of Chang cells was conditionally enhanced by HBx transfection.

摘要

目的

建立稳定表达乙型肝炎病毒X蛋白的Chang肝细胞系,并探讨乙型肝炎病毒X蛋白诱导其侵袭潜能的生物学行为。

方法

采用聚合酶链反应从完整的乙型肝炎病毒基因组中扩增HBx基因。然后将该基因亚克隆到真核表达载体pcDNA3.1中,构建pcDNA3.1-HBx质粒。利用脂质体介导的基因转染将质粒导入人肝细胞系Chang,并检测HBx基因的稳定表达。

结果

通过逆转录-聚合酶链反应从转染的Chang肝细胞中克隆出HBx基因,并经电泳证实。验证了稳定转染表达HBx且具有恶性特征的Chang细胞,并在生长曲线、克隆形成能力、伤口愈合能力、迁移和转移方面与对照细胞进行了比较。

结论

成功建立了稳定表达含HBx基因的人Chang肝细胞系。HBx转染可使Chang细胞的侵袭潜能有条件地增强。

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