ABSTRACT
Efforts to preserve β-cell mass in the preclinical stages of type 1 diabetes (T1D) are limited by few blood-derived biomarkers of β-cell destruction. Platelets are proposed to be the sources of blood-derived biomarkers for a variety of diseases, and they show distinct proteomic changes in T1D. Thus, studying these proteomic changes may provide us with biomarkers of β-cell destruction. This paper is the Chinese translation of " Exocytosis protein DOC2B as a biomarker of type 1 diabetes" , published on " The Journal of Clinical Endocrinology & Metabolism" in May 2018 [Aslamy A, Oh E, Ahn M, et al. J Clin Endocrinol Metab, 2018, 103(5): 1966-1976] after obtaining the copyright from the original journal. This study aimed to investigate the changes in the exocytosis protein double C2 domain protein-β (DOC2B) in platelets and islets from T1D humans, pre-diabetic NOD mice, and from T1D patients after islet transplantation. The results showed that the DOC2B protein abundances were substantially reduced in platelets of prediabetic NOD mouse and new-onset T1D patients, while platelet DOC2B levels were increased after islet transplantation in T1D patients. Thus, reduction of DOC2B is an early feature of T1D, and DOC2B abundance may serve as a valuable in vivo indicator of β-cell mass as well as an earlier biomarker of T1D. (Chin J Endocrinol Metab, 2018, 34: 615-622)
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.
ABSTRACT
Background and purpose:Studies have shown that DOC-2 could work as a potential tumor suppressor geue,and the role of DOC-2 in terms of the inhibition of cell growth and its mechanism remain unknown.Our paper is to investigate the effect and mechanism of DOC-2 expression on the tumorigenesis viability of ovarian cancer cell line HO-8910 from the aspects of clone efficiency,cell cycle and animal model test.Methods:Three cell lines were used including HO-8910,8910-P93(transfected with DOC-2 gene) and 8910-pcDNA3.1(transfected with the vector pcDNA3.1).Firstly,soft agar method was used to measure the clone efficiency.The cell cycle were analyzed by flow cytometer.The tumorigenesis viability was compared by athymic mouse test.Results:After being transfected with DOC-2 gene,the clone efficiency of 8910-P93 was markedly reduced.There was no difference between the 8910-pcDNA3.1 and HO-8910.G1 and G2 arrest were observed for 8910-P93.The athymic mouse test showed that the neoplasm derived from 8910-P93 was much smaller than that in the controls.Conclusions:DOC-2 could iniibit the tumorigenesis viability of human ovarian cancer line HO-8910.
ABSTRACT
Objective: To screen for proteins which can interact with phosphotyrosine-interacting domain (PID) of differentially expressed gene in human ovarian cancer cell line DOC-2 by yeast-two hybrid technique, so as to provide evidence for the signal pathway of DOC-2. Methods: The cDNA sequence of human DOC-2 gene was amplified and its PID domain (nDOC-2) was subcloned into the bait vector pGBKT7 of yeast two-hybrid system; the product was then used to screen an embryo brain cDNA library and the proteins interacting with nDOC-2 were identified. Quadrople dropout(QDO) medium and X-?-gal were used for selecting the positive clones. PCA was used to analyze the amplified sequence. After elimination of the false positive clones, the positive clones were sequenced and analyzed by bioinformatic methods. Results:Twenty-one candidate positive clones were obtained and 3 of them were plasmids encoding Homo sapiens partial mRNA for betaglycan (TBR III gene), Homo sapiens protocadherin gamma subfamily C 3 (PCDHGC3), and APLP1(amyloid beta precursor-like protein 1).Conclusion: The proteins obtained in this study may play important roles in the signal pathway of DOC-2, which provides a new orientation for DOC-2 gene therapy of ovarian cancers