Study on the expression and function of the mdr-1 gene transferred into hematopoietic cells of rabbit bone marrow in vitro
- VernacularTitle:多药耐药mdr-1基因体外转染兔骨髓造血细胞及其表达与功能的研究
- Author:
Yi WANG
- Publication Type:Journal Article
- Keywords:
Multidrug resistance gene;
Transfection;
Hematopoietic cells
- From:
Journal of Chongqing Medical University
1986;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the optimal transfection conditions in which the concentrated virus supernatant transfer the foreign mdr-1 gene into hematopoietic cells of rabbit bone marrow,and test the expression and function of the mdr-1 gene in cells.Methods:An amphotropic virus producer cell line,PA317-HaMDR1/A1 which contains a full-length cDNA of human mdr-1 gene,was screened with colchicine(90ng/ml),and the supernatant was collected and concentrated.The bone marrow cells of the New Zealand rabbits were gathered and the mononuclear cells which contain many hematopoietic cells were enriched.The hematopoietic cells were cocultivated with concentrated virus supernatant and interleukins.The transfection rate was determined by SP immunohistochemical methods;the integration of mdr-1 gene in rabbit mononuclear cells was tested by PCR method ,and the physiological function of mdr-1 gene was tested by DNR extrusion test.Results:After screening by colchicine,the expression of P-glycoprotein(P-gp) of PA317-HaMDR1/A1 was enhanced.The foreign mdr-1 gene was transferred into hematopoietic cells of rabbit bone marrow successfully,and the transduction rate of 2 days,4days and 6 days were 22%,37%,and 39% respectively.However,the state of the 4 days was the best.The transferred mdr-1 gene had its physiological function.Conclusion:The foreign mdr-1 gene can be transferred into hematopoietic cells of rabbit bone marrow successfully by concentrated virus supernatant transfection method,and the transferring gene can be expressed stably and effectively.The study has provided a basis for the further research on chemoprotection experiment of the mdr-1 gene transferred into the bone marrow cells.
