Development of new antitumor drugs and apoptotic signal transudation - Application to feline leukemia and canine lymphoma -

抗肿瘤新药开发及细胞凋亡信号转导-在猫白血病和犬淋巴瘤中的应用-

基本信息

批准号：
12460135
负责人：
KUWABARA Mikinori
金额：
$ 9.22万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12460135/
关键词：
LEUKEMIA LYMPHOMA ANTICANCER DRUG APOPTOSIS CASPASE CELL DEATH NECROSIS TUMOR CELL LINES

项目摘要

Clinically, the combination treatment of solid tumor cells with an anticancer drug and radiation was widely used to enhance cell killing. 1-(3-C-Ethynyl-β-D-ribo-pentofuranosyl) cytosine (ECyd) and 2-chrolo-deoxyadenosine (Cl-Ade) was newly developed as anticancer drugs to induce cell death by inhibiting RNA synthesis. In this study, we examined whether the exposure of these anticancer drugs or /and ionizing radiation to human gastric adenocarcinoma MKN45 cells, human lekemia cell line MOLT-4 cells induce apoptosis and canine lymphoma cell line CL-1. In MOLT-4 cells, Fas expression and caspase 8/3 activation apoptotic cell death was observed after treatment of Cl-Ade, followed by apoptotic cell death. 2Cl-Ade and ECyd showed to induce apoptotic cell death in canine lymphoma cell CL-1 cells. MKN45 cells were treated with 0.1 μM ECyd for 1 h before irradiation. After irradiation with 20 Gy, apoptotic as well as swelling cells were morphologically discriminated by fluorescence microscopy … More combined with propidium iodide staining or electron microscopy and were scored. When cells were treated with ECyd alone, only 5 % of either apoptotic or necrotic cells appeared. When cells were exposed to X rays alone, about 5 % of apoptotic and about 45 % of swelling cells appeared. However, when cells were exposed to X rays in the presence of 0.1 μM ECyd, about 25 % of total cells was apoptotic cells and about 10 % of total cells was swelling cells. Apoptosis induced by treating cells with ECyd and X irradiation was significantly reduced by the treatment with Ac-DEVD-CHO (caspase 3 inhibitor) or TPCK (chymotrypsin-like protease inhibitor).These results suggested that caspase 3 and chymotrypsin-like proteases were responsible for apoptosis induced by co-treatment with ECyd and X rays. In this study, it was demonstrated that co-treatment of MKN45 cells with ECyd and X rays activated G2-phase-linked apoptotic signaling associated with caspase 3 and chymotrypsin like protease. These data may provide useful information to develop clinical treatment of radiation combined with anticancer drugs for solid tumors. Less

临床上，实体瘤细胞与抗癌药物和放射线的联合治疗被广泛用于增强细胞杀伤作用，1-(3-C-乙炔基-β-D-核糖-呋喃糖基)胞嘧啶(ECyd)和2-氯脱氧腺苷。（Cl-Ade）是新开发的抗癌药物，通过抑制 RNA 合成来诱导细胞死亡。在这项研究中，我们检查了这些抗癌药物的暴露是否或/和。电离辐射对人胃腺癌MKN45细胞、人白血病细胞系MOLT-4细胞和犬淋巴瘤细胞系CL-1诱导细胞凋亡。在MOLT-4细胞中，观察到Fas表达和caspase 8/3激活后细胞凋亡。 Cl-Ade，随后是细胞凋亡，2Cl-Ade 和 ECyd 显示可诱导犬淋巴瘤细胞凋亡。 MKN45 细胞在照射前用 0.1 μM ECyd 处理 1 小时，用 20 Gy 照射后，通过荧光显微镜结合碘化丙啶染色或电子显微镜对凋亡和肿胀细胞进行形态区分并进行评分。当细胞单独用 ECyd 处理时，只有 5% 的细胞凋亡或坏死。当细胞单独暴露于 X 射线时，约 5% 出现凋亡，约 45% 出现肿胀细胞，然而，当细胞在 0.1 μM ECyd 存在下暴露于 X 射线时，约 25% 的细胞出现凋亡。凋亡细胞，约10%的细胞是肿胀细胞，用ECyd和X射线处理细胞诱导的细胞凋亡显着减少。 Ac-DEVD-CHO（半胱天冬酶 3 抑制剂）或 TPCK（胰凝乳蛋白酶样蛋白酶抑制剂）。这些结果表明，半胱天冬酶 3 和胰凝乳蛋白酶样蛋白酶是 ECyd 和 X 射线联合治疗诱导的细胞凋亡的原因。结果表明，ECyd 和 X 射线共同处理 MKN45 细胞可激活 G2 相相关的凋亡信号传导与caspase 3和糜蛋白酶样蛋白酶相比，这些数据可能为开发放射联合抗癌药物治疗实体瘤的临床治疗提供有用的信息。