Molecular biology and analysis of related genes of apoptosis in human gastric carcinomas and precancerous lesions

人胃癌及癌前病变细胞凋亡相关基因的分子生物学及分析

• 批准号: 07670204
• 负责人: ITO Hisao
• 金额: $ 1.54万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670204/
• 关键词: apoptosis; human gastric cancer; gastric adenoma; p53 gene; cell cycle; EB virus; anti cancer adents; Fas antigen; 制癌剤; EVウイルス; 増殖活性

1. in vivo analysis1) Apoptotic cells were extremely rare in normal gastric mucosa, whle a few apoptotic cells were present in deeper portions of the metaplastic glands. The number of apoptotic cells or apoptotic index (AI) was counted in 30 tubular adenomas. AI was higher in the tubular adenomas with severe atypia than in those with mild atypia. AI was analyzed in 15 surgically removed stomachs in which adenoma and carcinoma were found simultaneously. AI was 4.9+2.1 in the former and 3.9+1.1 in the latter, the value being significantly different (P<0.05). On the other hand, proliferative index (PI) was higher in the carcinomas.2) AI was 3.8+1.4 in 15 gastric cancers carrying wild type p53 and 4.9+1.2 in 15 cancers carrying mutated p53 gene, the value being significantly higher in the latter (P<0.05).3) AI was significantly lower in 17 EB-virus related gastric carcinomas with lymphoid stroma than other any histological type of gastric carcinomas (P<0.05).4) AI increased significantly t … More o 6.1+3.6 in 10 patients received continuous intravenous 5-FU at 500mg/day for 7 days preoperatively, compared with 10 patients without 5-FU (3.2+1.8 : P<0.05).2. in vitro analysis1) 1mM-5FU variably induced apoptosis in MKN-74 and MKN-45, both of which carries wild type p53 gene, but not MKN28 carrying mutated p53 gene and KATO-III p53 gene being deleted. Western blot analysis showed increasing of P53 and P21 in MKN-74 and MKN-45.2) Fas was detected to be expressed variably in the six human gastric carcinoma cell lines. The expression level was highest for MKN-74 and MKN-45 carrying wild type p53 gene. Anti-Fas antibody of 100ng/ml induced apoptosis, the value of AI being correlated with the expression levels of FAS antigen.3) Treatment with taxol of 1 uM arrested in G2/M,followed by apoptosis in the human gastric carcinoma cell lines. Susceptibility to taxol did not correlate with p53 gene status.These results overall indicate that gastric cancer cells might be variably induced apoptosis by a variety of stimuli, and signal transduction relating apoptosis partly depends on the p53 protein. Less

1. 1）正常胃粘膜中凋亡细胞极少，而在化生腺体的深部有少量凋亡细胞。对30例管状腺瘤进行凋亡细胞计数或凋亡指数（AI）测定。重度垂体腺瘤的AI高于轻度垂体腺瘤。对15例同时发现腺瘤和癌的手术切除胃标本进行了AI分析。AI前者为4.9 ± 2.1，后者为3.9 ± 1.1，差异有显著性（P<0.05）。（2）15例p53基因野生型胃癌的增殖指数（AI）为3.8 ± 1.4，15例p53基因突变型胃癌的AI为4.9 ± 1.2，后者的价值要高得多17例EB病毒相关性胃癌伴淋巴样间质组AI显著低于其他组织学类型胃癌（P <0.05）。 ...更多信息 o术前5-FU 500 mg/d持续静脉滴注7天的10例患者为6.1 ± 3.6，未使用5-FU的10例患者为3.2 ± 1.8，差异有统计学意义（P<0.05）.体外分析1）1 mM-5 FU可诱导携带野生型p53基因的MKN-74和MKN-45细胞凋亡，而不诱导携带突变型p53基因和缺失型KATO-Ⅲ p53基因的MKN-28细胞凋亡。Westernblot分析显示MKN-74和MKN-45细胞中P53和P21蛋白表达增加。携带野生型p53基因的MKN-74和MKN-45的表达水平最高。100 ng/ml的抗Fas抗体可诱导胃癌细胞凋亡，其凋亡指数与FAS抗原表达水平相关。3）1 μ M的紫杉醇可使胃癌细胞阻滞于G2/M期，并诱导胃癌细胞凋亡。p53基因状态与胃癌细胞对紫杉醇的敏感性无关，提示多种刺激均可诱导胃癌细胞凋亡，而与凋亡相关的信号转导部分依赖于p53蛋白。少

项目成果

Satoshi Ohfuji: "Low frequency of apoptosis in Epstein-Barr virusassociated gastric carcinoma with lymphoid stroma" International Journal of Cancer. 68. 1-7 (1996)

Satoshi Ohfuji：“Epstein-Barr 病毒相关胃癌与淋巴基质的细胞凋亡频率较低”国际癌症杂志。

Yoshihito Gomyo: "Numerical aberration and point mutation of p53 gene in human gastric intestinal metaplasia and well-differentiated adenocarcinoma : Analysis by fluorescence in situ hybridization (FISH) and PCR-SSCP" International Journal of Cancer. 66.

Yoshihito Gomyo：“人胃肠化生和高分化腺癌中 p53 基因的数值畸变和点突变：荧光原位杂交 (FISH) 和 PCR-SSCP 分析”国际癌症杂志。

Satoshi Ohfuji: "Low frequency apoptosis in Epstein-Barr virus-asociated gastric carcinoma with lymphoid stroma" International Journal of Cancer. 68. 1-7 (1996)

Satoshi Ohfuji：“Epstein-Barr 病毒相关胃癌与淋巴基质的低频细胞凋亡”国际癌症杂志。

Masato Ishida: "Apoptosis in human gastric mucosa, chronic gastritis, dysplasia and carcinoma : analysis by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick endlabelling" Virchows Archiv. 428. 229-235 (1996)

Masato Ishida：“人胃粘膜细胞凋亡、慢性胃炎、不典型增生和癌：通过末端脱氧核苷酸转移酶介导的 dUTP-生物素缺口末端标记进行分析” Virchows Archive。

Hisae Hayashi: "Expression of Fas antigen and its mediation apoptosis in human gastric cancer cell lines" Japanese Journal of Cancer Research. 88. 49-57 (1997)

Hisae Hayashi：“人胃癌细胞系中Fas抗原的表达及其介导的细胞凋亡”日本癌症研究杂志。