›› 2011, Vol. 31 ›› Issue (4): 387-.doi: 10.3969/j.issn.1674-8115.2011.04.001

• Original article (Basic research) •     Next Articles

Effects of anti-PNAS-2 single chain antibody on apoptosis of human leukemic U937 cells

XIAO Zhen-yu1, CHEN Fang-yuan1, WANG Hai-rong1, WU Ying-li2, ZHONG Ji-hua1, |ZHONG Hua1   

  1. 1.Department of Hematology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200127, China； 2.Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai |Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2011-04-28 Published:2011-04-28
  • Supported by:

    Research  Foundation  of  Renji  Hospital-Shanghai  Jiaotong  University  School  of  Medicine, ZD0704；National Natural Science  Foundation  of  China, 81000212

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Abstract:

Objective To observe specific binding ability of anti-PNAS-2 single chain antibody with PNAS-2 antigen and its effects on apoptosis of human leukemic U937 cells. Methods The pMIG-PNAS-2ScFv, anti-PNAS-2 single chain antibody expression vector, was transfected into U937 cell line by retroviral transfection method. Untransfected U937 cells and pMIG transfected with empty vector were used as controls. Transfection rate was measured by flow cytometry. Single chain antibody expression and ScFv-antigen specific binding were measured by laser confocal microscopy. Cell apoptosis was detected by flow cytometry and laser confocal microscopy. Expression of PNAS-2 and active caspase-3 was detected by Western blotting. Results Anti-PNAS-2 single chain antibody coding sequence was successfully transfected into U937 cells and its expressed single chain antibody could bind to PNAS-2 antigen with a high specificity. Compared with the control cells, pMIG-PNAS-2ScFv cells had a higher apoptosis rate (P<0.05) and increase in active caspase-3 expression，but no difference was found in PNAS-2 expression. Conclusion Anti-PNAS-2 single chain antibody can specifically bind PNAS-2 antigen and induce apoptosis of U937 cells, which may be related to the inactivation of PNAS-2 after closure of the functional site.

Key words: PNAS-2, single chain antibody, U937 cell, apoptosis