›› 2009, Vol. 27 ›› Issue (6): 3-466.

• 论著 • Previous Articles     Next Articles

Inhibition of Plasmodium yoelii Circumsporozoite Protein on the Activation of Nuclear Transcription Factor in Hepatoma Cells Stimulated by TNF-α

DING Yan,CHEN Ji-de,ZHOU Tao-li,FU Yong,PENG Xiao-hong,XU Wen-yue*   

  1. Department of Pathogenic Biology,The Third Military Medical University,Chongqing 400038,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-12-30 Published:2009-12-30

Abstract: Objective To investigate on the effect of Plasmodium yoelii(BY265 strain)circumsporozoite protein (CSP)on the activation of nuclear transcription factor κB(NF-κB)in hepatoma cells(HepG2)stimulated by TNF-α. Methods Entire coding sequence of CSP was reverse transcribed and amplified by RT-PCR with sporozoite total RNA as template,then cloned into pFLAG-CMV8 for construction of the recombinant plasmid pFLAG-CMV8-CSP. Indirect immunofluorenscence staining with rabbit anti-csp was applied to verify the expression and distribution of FLAG-CSP fusion protein in HepG2. Three groups were established for the experiment: group A with HepG2 transfected by pFLAG-CMV8 as negative control, group B with HepG2 transfected by pFLAG-CMV8 and stimulated by 100 ng/ml TNF-α, and group C with HepG2 transfected by pFLAG-CMV8-CSP and stimulated by 100 ng/ml TNF-α. Dual-luciferase assay and EMSA were performed to detect the nuclear translocation and activation of NF-κB, to observe if pFLAG-CMV8-CSP suppressed the activation of NF-κB in HepG2 stimulated by TNF-α. Result The expression of pFLAG-CMV8-CSP was localized on cytoplasm of HepG2. The activity ratio of firefly luciferase to Renilla luciferase in group C(0.228±0.029) was significantly lower than both groups A(0.438±0.085)and B(0.571±0.030) (P<0.05). EMSA showed that the band in group C was significantly weaker than group B. Conclusion Plasmodium yoelii CSP localizes in the cytoplasm of HepG2 and inhibits the activation and nuclear translocation of NF-κB in HepG2 stimulated by TNF-α.

Key words: Plasmodium yoelii, Circumsporozoite protein, Nuclear transcription factor κB, Hepatoma cell