STRATEGIC DOWN-REGULATION OF DNA-POLYMERASE-BETA BY ANTISENSE RNA SENSITIZES MAMMALIAN-CELLS TO SPECIFIC DNA-DAMAGING AGENTS
STRATEGIC DOWN-REGULATION OF DNA-POLYMERASE-BETA BY ANTISENSE RNA SENSITIZES MAMMALIAN-CELLS TO SPECIFIC DNA-DAMAGING AGENTS
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DOI：
10.1093/nar/23.19.3810
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发表时间：
1995-10-11
影响因子：
14.9
通讯作者：
WILSON, SH
中科院分区：
文献类型：
--
作者：
HORTON, JK;SRIVASTAVA, DK;WILSON, SH
先前，小鼠NIH 3 T3细胞用人DNA聚合酶β（β-pol）cDNA以反义方向并在金属硫蛋白启动子的控制下稳定转染[Zmudzka，B.Z. Wilson，S. H.（1990）Som.细胞分子将军，16，311-320]。为了评估通过反义方法增强化疗疗效的可行性并确认β-pol在细胞DNA修复中的作用，我们在反义细胞系中β-pol下调的条件下寻找对DNA损伤剂的敏感性增加。只有当β-pol在DNA修复途径中具有限速作用时，才能预期这种致敏作用。测试了一些试剂：顺式二氨二氯铂II（顺铂）; 1，3-双（2-氯乙基）-1-亚硝基脲（BCNU）;电离辐射和放射模拟药物博来霉素;双功能烷化剂氮芥和L-苯丙氨酸芥（美法仑）;单功能烷化剂甲磺酸甲酯（MMS）和紫外线（UV）辐射。在顺铂和紫外线照射的情况下，观察到细胞毒性的显着增强。这两种药物造成的损伤被认为是通过核苷酸切除修复（NER）途径修复的。结果表明，在该细胞系中，β-pol参与并限制NER的速率。我们建议通过反义方法下调β-pol可用于增强顺铂和其他DNA损伤化疗药物的细胞毒性作用。
Previously, mouse NIH 3T3 cells were stably transfected with human DNA polymerase beta (beta-pol) cDNA in the antisense orientation and under the control of a metallothionein promoter [Zmudzka,B.Z. and Wilson,S.H. (1990) Som. Cell Mol. Gen., 16, 311-320]. To assess the feasibility of enhancing the efficacy of chemotherapy by an antisense approach and to confirm a role for beta-pol in cellular DNA repair, we looked for increased sensitivity to DNA damaging agents under conditions where beta-pol is down-regulated in the antisense cell line. Such a sensitization is anticipated only where beta-pol is rate-limiting in a DNA repair pathway. A number of agents were tested: cis-diamminedichloroplatinum II (cisplatin); 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU); ionizing radiation and the radio-mimetic drug bleomycin; the bifunctional alkylating agents nitrogen mustard and L-phenylalanine mustard (melphalan); the monofunctional alkylating agent methyl methane sulfonate (MMS) and ultraviolet (UV) radiation. In the cases of cisplatin and UV radiation, a significant enhancement of cytotoxicity was observed. Damage as a result of both of these agents is thought to be repaired by the nucleotide excision repair (NER) pathway. The results suggest that, in this cell line, beta-pol is involved in and is rate-limiting in NER. We propose that down-regulation of beta-pol by antisense approaches might be used to enhance the cytotoxic effects of cisplatin and other DNA damaging chemotherapeutic agents.