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

在临床上，实体瘤细胞与抗癌药物和辐射的联合处理被广泛用于增强细胞杀伤。 1-（3-C-乙基-β-D-核硫素基）胞嘧啶（ECYD）和2-Chrolo-Deoxydenosine（Cl-Dade）是新开发的，是作为抗癌药物通过抑制RNA合成诱导细胞死亡的抗癌药物。在这项研究中，我们检查了这些抗癌药物的暴露是 /和 /和电离辐射是否会暴露于人类胃腺癌MKN45细胞，人Lekemia细胞系MOLT-4细胞诱导凋亡和犬淋巴瘤细胞系CL-1。在MOLT-4细胞中，FAS表达和caspase 8/3激活凋亡细胞死亡在治疗CL-DADE后，然后观察到凋亡细胞死亡。 2Cl-dade和Ecyd表明在犬淋巴瘤细胞CL-1细胞中诱导凋亡细胞死亡。辐照前，将MKN45细胞用0.1μMECYD处理1小时。用20 Gy辐照后，通过荧光显微镜在形态上歧视凋亡和肿胀细胞……更多地与碘化丙啶染色或电子显微镜结合并进行了评分。当单独用ecyd处理细胞时，只有5％的凋亡或坏死细胞出现。当细胞单独暴露于X射线时，出现了约5％的凋亡和约45％的肿胀细胞。但是，当细胞在存在0.1μMECYD的情况下暴露于X射线时，约有25％的总细胞是凋亡细胞，约10％的总细胞是肿胀的细胞。通过AC-DEVD-CHO（caspase 3抑制剂）或TPCK（chymotrypsin-like蛋白酶抑制剂）处理，用ECYD和X辐照细胞诱导的细胞诱导的凋亡显着降低。这些结果表明，胱天蛋白酶3和胰凝乳蛋白酶样蛋白酶是与ECYD和X射线共处引起的凋亡。在这项研究中，证明了MKN45细胞与ECYD和X射线的共同处理激活了与caspase 3和chymotrypsin（如蛋白酶）相关的G2相连接的凋亡信号传导。这些数据可能会提供有用的信息，以开发辐射的临床治疗以及抗癌药物的实体瘤。较少的