Expression of recombined human endothelial monocyte-activating polypeptideⅡand determination of its activity
- VernacularTitle:人内皮-单核活化多肽-2的重组表达及其活性鉴定
- Author:
Yun GAO
;
Liang-Hua WANG
;
Na REN
;
Ming-Juan SUN
;
Ai-Yun GUO
;
Bing-Hua JIAO
;
- Publication Type:Journal Article
- Keywords:
endothelail monocyte-activating polypeptideⅡ;
cloning and expression;
antitumor effect;
suppression proliferation;
apoptosis
- From:
Chinese Journal of Cancer Biotherapy
1995;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To chine and express the recombinant human endothelial monocyte-activating polypeptide-Ⅱ(EMAP-Ⅱ)and identify its anti-tumor biological activities.Methods: EMAP-Ⅱ_(147-312)was expressed by the expression vector pMAL-p2x and E.coli BL-21 and the product was purified.The production of tissue factor(TF)in human umbili- cal vein endothelial cell ECV-304 mediated by the recombinant EMAP-Ⅱwas determined by chemiluminescence sub- strate.The promoting effect of recombinant EMAP-Ⅱon TNF?-induced ECV-304 cell.Apoptosis was determined by flow cytometry.Its inhibitory effect on human pancreaic cancer cell SW1990 proliferation was determined by MTT method. Results:DNA sequencing verified that EMAP-Ⅱwas correctly cloned.The molecular mass of the protein identified by SDS-PAGE was consistent with the theoretic value.The productivity of recombinant EMAP-Ⅱwas 500?g per 1 g bacteria (wet mass).The purified product induced expression of tissue factor(TF)in ECV-304 cells;it also enhanced the sensi- tivity of ECV-304 cells to the apoptotic effect of TNF?([16.6?2.5]% vs[25.6?2.3]%,P
