突变型二氢叶酸还原酶基因对人外周血CD34+细胞抗甲氨蝶呤的保护作用
[Protective effect of mutant dihydrofolate reductase gene on human peripheral blood CD34+ cells against methotrexate].
作者信息
Wu R, Yang Z, Liu J
机构信息
Cancer Institute(Hospital), Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100021.
出版信息
Zhonghua Zhong Liu Za Zhi. 1999 May;21(3):186-9.
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.
目的
研究转导突变型二氢叶酸还原酶（mDHFR）基因的人外周血造血祖细胞是否能增强其对甲氨蝶呤（MTX）的耐药性。
方法
用高梯度磁性细胞分选系统（MACS）富集后，将外周血CD34+细胞用含人mDHFR cDNA的逆转录病毒上清液转染。利用造血祖细胞集落形成来评估mDHFR赋予的对MTX的耐药性。计算每接种500个细胞形成的集落数，并以集落形成百分比表示。
结果
在含20 nmol·L-1 MTX的培养基中体外筛选14天后，转导mD-HFR的CD34+细胞和空载体转导的CD34+细胞的集落形成百分比分别为29.4%和16.3%（P<0.01）。对MTX的耐药性提高了两倍。
结论
转导mDHFR基因可赋予人造血祖细胞对MTX毒性的耐药性。
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