EFFECT OF CALMODULIN INHIBITORS ON ADRIAMYCIN RESISTANCE

这项建议的总体目标是评估 体内钙调蛋白抑制剂对阿霉素抗肿瘤活性的影响 渐进性耐药细胞与阿霉素对正常人的毒性 易受感染的目标组织。这些研究将对扩大我们的 理解阻力的机制，并最终在设计中 治疗耐药肿瘤的化疗策略 蒽环类药物化疗。这项研究中使用的肿瘤系统 将包括对父母敏感和对阿霉素逐渐耐药的 P388、L1210小鼠白血病和B16-BL6小鼠的变异体 黑色素瘤。需要评估的钙调蛋白抑制剂包括 N-(4-aminobutyl)-5-chloro-2-naphtalenesulfonamide(W-13)、三氟拉嗪、 氯丙嗪和氯丙嗪。耸人听闻，循序渐进 用于体内抗肿瘤研究的耐药肿瘤最初将 其特征是确定钙调蛋白抑制剂对血管内皮细胞 阿霉素的蓄积、滞留和细胞毒性作用。蜂窝 阿霉素水平将通过荧光法和激光流动法进行测量 流式细胞术检测细胞内分布的异质性 阿霉素荧光。药物诱导的细胞毒性效应将基于 软琼脂集落形成率。 肿瘤细胞的增殖性变化将通过测量 ~3H-胸腺嘧啶核苷的摄取和细胞内微扰的测定 用流式细胞仪进行周期遍历。体内的抗肿瘤作用将是 通过测量肿瘤体积的变化和肿瘤的延长来确定 经过处理的动物的寿命。阿霉素诱导的可能恶化 钙调蛋白抑制剂对正常靶组织的体内毒性 将基于评估十二指肠隐窝的增殖和损伤以及 骨髓细胞和心肌损伤的定量检测。

The overall objective of this proposal is to evaluate the effect of calmodulin inhibitors in vivo, on the antitumor activity of adriamycin in progressively resistant cells and the toxicity of adriamycin to normal susceptible target tissues. These studies will be important to expand our understanding of the mechanisms of resistance and ultimately in the design of chemotherapeutic strategies for the management of tumors resistant to anthracycline chemotherapy. The tumor systems to be used in this study will include the parent-sensitive and progressively adriamycin-resistant variants of the P388 and L1210 mouse leukemia and the B16-BL6 mouse melanoma. The calmodulin inhibitors to be evaluated are N-(4-aminobutyl)-5-chloro-2-naphtalenesulfonamide (W-13), trifluoperazine, prochlorperazine and chlorpromazine. The sensitve and progressively resistant tumors for in vivo antitumor studies will be initially characterized to determine the effect of calmodulin inhibitors on the accumulation, retention, and cytotoxic effects of adriamycin. Cellular adriamycin levels will be measured by fluorometry and also by laser flow cytometry to determine heterogeneity in the cellular distribution of adriamycin fluorescence. Drug-induced cytotoxic effects will be based on the colony-forming ability of the treated tumor cells in soft agar. Proliferative changes in the tumor cells will be determined by measuring the uptake of 3H-thymidine and the determination of perturbations in cell cycle traverse by flow cytometry. Antitumor effects in vivo will be determined by measuring changes in tumor volume and the prolongation in life-span of treated animals. Possible exacerbation in adriamycin-induced toxicity in vivo to normal target tissues due to the calmodulin inhibitors will be based on evaluating proliferation and damage in duodenal crypt and bone marrow cells and by quantitating myocardial damage.