APPROARCH AT MOLECULAR LEVEL FOR MECHANISMS OF MULTIDRUG-RESISTANCE AND ENHANCEMENT OF ANTICANCER DRUGS WITH HYPERTHERMIA

分子水平的多药耐药机制和热疗增强抗癌药物的研究

• 批准号: 11670890
• 负责人: KAWASAKI Shoji
• 金额: $ 2.05万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670890/
• 关键词: adriamycin; multidrug resistance; hyperthermia; Ehrlich ascites tumor cells; anticancer drugs; ion channel; MGMT; alkylating agent; WAF1; wortmannin; 分化度; 喉頭癌; アントシアニジン; Caイオン; Calcein

It is an important problem that tumor cells treated with anticancer drugs expressed multidrug resistance. Therfore, resistant mechanisms of varius mammalian cells were studied, MDR1 gene concerning resistance to adriamycin were appeared in our established cells of Ehrlich ascites tumor. Its resistance related to eflux pump of p-gp which is in cellular membrane, and the penetration of adriamycin into cells was affected by both of ion channels of Na^+/H^+ and Cl^-/HCO_3. Heating showed marked enhancement of cell killing effects of adriamycin. Abtitumor effects of adriamycin were tested on in vivo with or without treatment of mild hyperthermia. However, antitumor effects were shown not so much. On the other hand, efflux of adriamycin was inhibited with cepharanthine, that is, cepharanthine enhanced a killing effect of adriamycin. Antitumor effects of adriamycin were affected with intraa- and extracellure pH. Intracellular accumulation of adriamycin was small amount at acidic condition. It was depended to slow infflux of adriamycin. It is concluded that antitumor effects of adriamycin were not so large enhancement with hyperthermia as it is low pH condition in tumor and hyperthermia induced low pH more than without treatment. On the othe hand, those cells showed the other unknown resistant mechanisims to adriamycin. It is indicated that adriamysin-resistant cells of Ehrlich ascites tumor have at least two expression processes of resistance to adriamycin. There are many mechanisms on resistant to anticancer drugs. Hyperthermia enhanced markedly killing effects of alkylating agents in vitro and in vivo. The sensitivity to alkylating agents related to 06-methylguanine-DNA methyltransferase(MGMT). As MGMT expression was depressed by methylation, it is investigated that relationship between level of methylation and killing effect of alkylating agent in several human squarmous carcinoma cells. Methylation of MGMT is closely related to the sennsitivity for alkylating agents.

抗癌药物治疗的肿瘤细胞表现出多药耐药性是一个重要问题。因此，对多种哺乳动物细胞的耐药机制进行了研究，在我们建立的艾氏腹水瘤细胞中发现了与阿霉素耐药相关的MDR1基因。其阻力与细胞膜p-gp的外排泵有关，阿霉素向细胞内的渗透受Na^+/H^+和Cl^-/HCO_3离子通道的影响。加热显示阿霉素的细胞杀伤作用显着增强。在有或没有轻度热疗治疗的情况下体内测试了阿霉素的抗肿瘤作用。然而，抗肿瘤作用并没有表现出那么多。另一方面，千金藤素抑制了阿霉素的外流，即千金藤素增强了阿霉素的杀伤作用。阿霉素的抗肿瘤作用受到细胞内和细胞外 pH 值的影响。酸性条件下阿霉素在细胞内蓄积量较小。它可以减缓阿霉素的流入。结论是，热疗后阿霉素的抗肿瘤作用并没有那么大的增强，因为肿瘤内处于低pH条件，并且与未治疗相比，热疗引起的低pH更多。另一方面，这些细胞对阿霉素表现出其他未知的耐药机制。表明艾利希腹水瘤阿霉素耐药细胞至少存在两种​​对阿霉素耐药的表达过程。抗癌药物耐药的机制有很多。高温可显着增强烷化剂的体外和体内杀伤作用。对烷化剂的敏感性与06-甲基鸟嘌呤-DNA甲基转移酶(MGMT)有关。由于MGMT表达被甲基化抑制，因此研究了几种人鳞癌细胞中甲基化水平与烷化剂杀伤作用之间的关系。 MGMT的甲基化与烷化剂的敏感性密切相关。

项目成果

Asaumi J, Matsuzaki H, Kawasaki S, Kuroda M, Takeda Y, Kishi K, and Hiraki Y: "Influence of bovine calf serum on the intracellular accumulation and retention of adriamycin"Anticancer Research. 20 (2A). 769-772 (2000)

Asaumi J、Matsuzaki H、Kawasaki S、Kuroda M、Takeda Y、Kishi K 和 Hiraki Y：“小牛血清对阿霉素细胞内积累和保留的影响”抗癌研究。

Asaumi J, Higuchi Y, Matsuzaki H, Murakami J, Kawasaki S, Kuroda M, Shibuya K, Konouchi H, Hisatomi M, Wakasa T, Kishi K, and Hiraki Y: "Thermochemotherapy of a human sailvary adenocarcmoma cell line."Oncol Rep.. 9 (2). 365-369 (2002)

Asaumi J、Higuchi Y、Matsuzaki H、Murakami J、Kawasaki S、Kuroda M、Shibuya K、Konouchi H、Hisatomi M、Wakasa T、Kishi K 和 Hiraki Y：“人帆腺癌细胞系的热化疗。”Oncol Rep