Xu L F, Ge K, Zheng Z C, Sun L Y, Liu X Y
Shanghai Institute of Biochemistry, Academia Sinica.
Shi Yan Sheng Wu Xue Bao. 1996 Dec;29(4):385-93.
A negative selection system for glioma gene therapy was established in vitro. C 6 rat glioma cells were infected with recombined retrovirus which contain Escherichia coli cytosine deaminase (EC-CD) gene. The enzyme CD can transform the non-toxic prodrug 5-Fluorocytosine (5-FC) to the highly cellular toxic compound 5-Fluorouracil (5-FU). The growth inhibition studies proved that CD-positive cells were highly sensitive to 5-FC, the IC50 about 3 mumol/L, compared with an IC50 of approximately 6000 mumol/L in parental C 6 cells. Both CD-positive and negative cells were sensitive to 5-FU at very low concentration (IC50 < 1 mumol/L). Mixed cellular assay showed CD-positive cells had "bystander effect" on CD-negative cells when exposed to 5-FC. Our results demonstrate that EC-CD gene should be an efficient suicide gene for the treatment of glioma.
在体外建立了一种用于胶质瘤基因治疗的负性选择系统。用含有大肠杆菌胞嘧啶脱氨酶(EC-CD)基因的重组逆转录病毒感染C6大鼠胶质瘤细胞。胞嘧啶脱氨酶(CD)可将无毒的前体药物5-氟胞嘧啶(5-FC)转化为细胞毒性很强的化合物5-氟尿嘧啶(5-FU)。生长抑制研究证明,CD阳性细胞对5-FC高度敏感,IC50约为3μmol/L,而亲代C6细胞的IC50约为6000μmol/L。CD阳性和阴性细胞在极低浓度(IC50<1μmol/L)时对5-FU均敏感。混合细胞试验表明,当暴露于5-FC时,CD阳性细胞对CD阴性细胞有“旁观者效应”。我们的结果表明,EC-CD基因应是治疗胶质瘤的一种有效的自杀基因。