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Development of antitumor drugs to enhance apoptotic cell death and clinical application

增强细胞凋亡的抗肿瘤药物的研制及临床应用

基本信息

批准号：
12660266
负责人：
INANAMI Osamu
金额：
$ 2.62万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
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/grant/KAKENHI-PROJECT-12660266/
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-rijbo-pentofuranosyl) cytosine (ECyd) was newly developed as an anticancer drug to induce cell death by inhibiting RNA synthesis. In this study, we examined whether the exposure of human gastric adenocarcinoma MKN45 cells to X rays in the presence of ECyd at the limited concentration ranges with no apoptosis induced apoptotic cell death. MKN45 cells were treated with 0.1μM ECyd for 1 h before irradiation. After irradiation with 20 Gy, apoptotic as well as swelling cells were morphologically discriminated by fluorescence microscopy 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 0.1μM ECyd, about 25% of totali 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.

临床广泛采用抗癌药物与放射联合治疗实体瘤细胞来增强细胞杀伤。1-（3- c -乙基-β- d -rijbo-戊呋喃基）胞嘧啶（ECyd）是一种新发现的通过抑制RNA合成诱导细胞死亡的抗癌药物。在本研究中，我们检测了人胃腺癌MKN45细胞暴露在ECyd存在的X射线下，在有限浓度范围内无凋亡是否会诱导凋亡细胞死亡。0.1μM ECyd处理MKN45细胞1 h。20 Gy辐照后，采用荧光显微镜联合碘化丙啶染色或电镜对凋亡细胞和肿胀细胞进行形态学区分并评分。单用ECyd处理细胞时，仅有5%的细胞出现凋亡或坏死。当细胞单独暴露在X射线下时，出现约5%的凋亡细胞和约45%的肿胀细胞。然而，当细胞暴露在0.1μM ECyd的X射线下时，约25%的细胞为凋亡细胞，约10%的细胞为肿胀细胞。用Ac-DEVD-CHO （caspase 3抑制剂）或TPCK （chymotrypsin样蛋白酶抑制剂）处理ECyd和X照射诱导的细胞凋亡明显减少。上述结果提示，ECyd和X射线共同作用诱导的细胞凋亡与caspase 3和凝乳胰蛋白酶样蛋白酶有关。本研究证明，ECyd和X射线共同处理MKN45细胞激活了与caspase 3和糜凝胰蛋白酶样蛋白酶相关的g2期凋亡信号。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。