Biol. Pharm. Bull., 32(4),724-727, April 2009

Notes

Heterozygous Disruption of the DNA Topoisomerase I Gene Confers Cellular Resistance to Camptothecin in Human Cells

Eriko TOYODA,^a,b Aya KUROSAWA,^a Michihiko FUJII,^a and Noritaka ADACHI^*,a

^a International Graduate School of Arts and Sciences, Yokohama City University; 22-2 Seto, Kanazawa-ku, Yokohama 236-0027, Japan: and ^b Pharmaceutical Research Laboratories, Nippon Kayaku Co., Ltd.; 3-31-12 Shimo, Kita-ku, Tokyo 115-8588, Japan. * To whom correspondence should be addressed. e-mail: nadachi@yokohama-cu.ac.jp

DNA topoisomerase I (Top1) is a ubiquitous nuclear enzyme that plays essential roles in various cellular processes, such as transcription or replication. Agents that target Top1, involving camptothecin and its derivatives, are among the most effective anticancer drugs used in the clinic. Previous work has suggested that the level of Top1 expression correlates with the cytotoxicity of camptothecin, but no direct evidence has been provided thus far in the context of human cells with a strictly isogenic genetic background. In this study, we perform heterozygous disruption of the Top1 gene (TOP1) by gene targeting in a human pre-B cell line, Nalm-6, which is karyotypically stable and normal for p53 status. We show that the heterozygous loss of the TOP1 gene does confer cellular resistance to camptothecin, to an extent comparable to that observed in the absence of functional p53 protein. Such a tolerance was not observed with other agents that target DNA topoisomerase II. Our results provide direct evidence that human cells with decreased Top1 levels are significantly more resistant to killing by camptothecin than are otherwise isogenic cells.

Key words DNA topoisomerase I; camptothecin; p53; gene targeting; Nalm-6