ABSTRACT
Objective To investigate the effect of APE1 on differentiation of peripheral blood mononuclear cells into osteoclast-like cells(OCL) which induced by macrophage colony stimulating factor (RANKL) and macrophage colony stimulating factor (M-CSF) .Methods Human peripheral blood mononuclear cells (PBMCs) were collected by density gradient separation ;Constructed APE1 siRNA expression vector Ad5v-APE1 siRNA was used to transfect PBMCs .Tartrate-resistant acid phosphatase (TRAP) method was conducted to identify the cells ,the expression level of APE1 was detected by Western blot ,the mRNA expression levels of Cathepsin K(CK) and V-ATPase were detected by RT-PCR .Results PBMCs transfected with APE1 siRNA had significantly lower protein expression of APE1 than untransfected cells (P< 0 .05) ;PBMCs could differentiate into OCL under the stimulation of RANKL and M-CSF ,the mRNA expression levels of CK and V-ATPase increased ;After APE1 siRNA treatment ,the number of OCL was reduced and the levels of CK and V-ATPase mRNA decreased .Conclusion PBMCs can differentiate into a large number of OCL induced by RANKL and M-CSF ,APE1 siRNA significantly inhibited differentiation of PBMCs into osteoclast-like cells , APE1 may be involved in the regulation of osteoclast-like differentiation process .
ABSTRACT
Apurinic/apyrimidinic endonuclease/redox factor 1(APE/Ref 1) is a multifunctional protein that is responsible for repairing DNA damage,it also infleunce reduction oxidation (redox) reaction and modulate the DNA binding activity of transcription factors,it is important to cell survival.Recently, much data concerning APE1/Ref 1 and information on APE1/Ref 1 in a wide variety of functions and systems have rapid growth,including its gene,function,distribution and relations with some diseases,especially nervous system diseases and tumors.