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The role of NF-κB in hepatocellular carcinoma cell

王剑虹 黄庆科 陈民新

中华医学杂志(英文版)2003,Vol.116Issue(5):747-752,6.
中华医学杂志(英文版)2003,Vol.116Issue(5):747-752,6.

The role of NF-κB in hepatocellular carcinoma cell

The role of NF-κB in hepatocellular carcinoma cell

王剑虹 1黄庆科 1陈民新1

作者信息

  • 1. Department of Gastroenterology, First Affiliated Hospital, Wenzhou Medical College, Wenzhou 325000, China
  • 折叠

摘要

Abstract

Objective To evaluate the role of nuclear factor-kappaB (NF-κB) and IκBα in hepatocellular cacinoma (HCC) SMMC7721 cells, the consequence of NF-κB inhibition in SMMC7721 cells transfected with mutated IκBα (mIκBα) plasmid and the effect of stable inhibition of NF-κB activity in combination with Doxorubicin.Methods Western blot was used to determine the expression of NF-κB and IκBα in SMMC7721 cells and normal liver cells. Nuclear protein was used to evaluate the binding of the 32 P-labeled tandem κB sequence using electrophoretic mobility shift assay and the expression of NF-κB using Western blot between SMMC7721 cells transfected with mIκBα plasmid (SMMC7721-MT) and control cells. Furthermore, cell viability was plotted between SMMC7721-MT and control cells. The binding of κB sequence and cell viability between SMMC7721-MT and control cells at different concentrations of Doxorubicin were also investigated.Results Western blot analysis for nuclear extract showed more P50 (NF-κB1) and P65 (RelA) expression in SMMC7721 cells compared with normal liver cells. The expression of cytosolic IκBα protein in SMMC7721 cells was less than that in normal cells. SMMC7721-MT cells inhibited NF-κB nuclear translocation at 0, 24, 48 and 96 hours. Furthermore, NF-κB cannot be detected in the nuclear protein of SMMC7721-MT cells by Western blot. By calculating cell viability, the proliferation of SMMC7721-MT cells was shown to be suppressed more significantly than that of control cells. NF-κB in untransfected cells was activated by Doxorubicin in a dose-dependent manner, but that in SMMC7721-MT cells was not induced at low concentrations of Doxorubicin. Compared with untransfected cells, the viability of SMMC7721-MT cells was significantly suppressed at the same concentration of Doxorubicin (P<0.01).Conclusions The present study demonstrates that upregulation of NF-κB and downregulation of inhibitory kappaB (IκBα) in SMMC7721 cells are related with the growth of hepatocellular cacinoma cells. Stable expression of mIκBα in SMMC7721-MT cells can inhibit NF-κB nuclear translocation and suppress cell growth. Furthermore, stable inhibition of NF-κB activity in combination with Doxorubicin can significantly inhibit cell proliferation in SMMC7721-MT cells. Thus, modulation of NF-κB may represent an improvement in the efficacy of HCC therapies and be worthy of further research and investigation.

关键词

hepatocellular cacinoma/nuclear factor κB/inhibitory κB alpha/Doxorubicin

Key words

hepatocellular cacinoma/nuclear factor κB/inhibitory κB alpha/Doxorubicin

分类

医药卫生

引用本文复制引用

王剑虹,黄庆科,陈民新..The role of NF-κB in hepatocellular carcinoma cell[J].中华医学杂志(英文版),2003,116(5):747-752,6.

中华医学杂志(英文版)

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