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目的研究转染耐药基因后对造血干细胞的保护作用方法以脂质体为载体,把二氢叶酸还原酶(DHFR)突变的对氨甲喋呤(MTX)耐药的基因转染到小鼠造血干细胞,再输回至小鼠体内后,对小鼠进行为期9周的大剂量MTX化疗。结果转染MTX耐药基因组小鼠化疗前后的粒细胞总数及体重均较稳定,而未转染基因组小鼠的粒细胞总数及体重呈明显下降(P<0.05)。小鼠存活率在转染基因组较对照组有明显提高。从小鼠脾脏提取DNA,经PCR扩增后,证实转染基因组有耐药基因的高效表达。结论此实验为肿瘤患者在大剂量化疗时,进行保护造血功能,提高化疗疗效的实验研究和临床应用,提供有参考价值的资料。
Objective To study the protective effect of drug-resistant gene on hematopoietic stem cells.Methods Lipofectamine 2000 was used to transfect the methotrexate (MTX) -resistant gene of dihydrofolate reductase (DHFR) into mouse hematopoietic stem cells, After being transfused back into the mice, the mice were given a high dose of MTX chemotherapy for 9 weeks. Results The total number and weight of granulocytes in the transfected MTX-resistant mice were stable before and after chemotherapy, while the number and weight of granulocytes in untransfected mice decreased significantly (P <0.05). The survival rate of mice in transfection of the genome significantly increased compared with the control group. DNA was extracted from the spleen of mice. After PCR amplification, it was confirmed that the transfected genome had high expression of resistant gene. Conclusions This experiment provides experimental data and clinical application for the cancer patients to protect the hematopoietic function and improve the curative effect of chemotherapy when using large doses of chemotherapy.