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[Protective effect of mutant dihydrofolate reductase gene on human peripheral blood CD34+ cells against methotrexate]
1Cancer Institute(Hospital), Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100021.
Objective:
To examine if human peripheral blood hematopoietic progenitor cells transduced with mutant dihydrofolate reductase(mDHFR) gene may increase their resistance to metrotrexate(MTX).
Methods:
After enrichment with a high-gradient magnetic cell sorting system(MACS), the peripheral blood CD34+ cells were transfected with supernatant of retrovirus containing human mDHFR cDNA. Colony formation of hematopoietic progenitor cells was utilized to evaluate mDHFR-conferred resistance to MTX. The number of colonies formed from every 500 cells seeded was counted and expressed as the percentage of colony formation.
Results:
After the mD-HFR-transduced CD34+ cells had been screened in vitro in medium containing 20 nmol.L-1 MTX for 14 days, the percentage of colony formation of mDHFR cDNA-transduced and of mock-transduced CD34+ cells was 29.4% and 16.3%, respectively (P < 0.01). The resistance to MTX was increased twofold.
Conclusion:
Transduction of mDHFR gene could confer the resistance of human hematopoietic progenitor cells to MTX toxicity.