Screening of DOC-2 interactive proteins and identification of the interaction between DOC-2 and TGF?Ⅲ receptor / 中国癌症杂志

ABSTRACT

Background and purpose:DOC-2 serves as one of the tumor suppressing genes in human ovarian cancer and plays a role in the process of cell growth and differentiation.This study was to investigate the role of DOC-2 in the TGF? signal pathway and verification of the interaction between DOC-2 and TGF?Ⅲ receptor.Methods:The bait vector was constructed by inserting the PID domain of DOC-2(nDOC-2)into yeast express vector pGBKT7.pGBKT7-nDOC2 was transformed into the yeast AH109 and confirmed to be expressed.After the human foetus brain cDNA library had been transformed,the positive clones was screened by both nutrition defect medium and X-?-gal.The putative positive clones were sequenced and analyzed to get the DOC-2 interactive proteins.Furthermore,after the DOC-2 cDNA and TGF?Ⅲ receptor cDNA had been co-transfected into the human ovarian cancer cell line HO-8910 together,the interaction between DOC-2 and TGF?Ⅲ receptor was investigated by immunoprecipitation and Western blot.Results:21 putative positive clones were picked after being screened and sequenced.Three of them were identified as Homo sapiens partial mRNA for betaglycan(TBR Ⅲ gene),homo sapiens protocadherin gamma subfamily C3(PCDHGC3)and APLP1(amyloid beta precursor-like protein 1).The analysis by immunoprecipitation and Western blot showed that the interaction between DOC-2 and TGF?Ⅲ receptor could form protein complex.Conclusions:The three encoding proteins might participate in the DOC-2 signal pathway.DOC-2 might play as an essential role in the TGF?signal pathway by interacting with TGF?Ⅲ receptor.