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induction of Apoptosis by L-myc Gene and p53 Gene.

L-myc基因和p53基因诱导细胞凋亡。

基本信息

批准号：
06671328
负责人：
SHIBA Mitsutoshi
金额：
$ 1.34万
依托单位：
Chiba University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06671328/
关键词：
p53 gene Apoptosis myc gene Non-small cell lung cancer PCR-SSCP Immunohistochemistry Ki-67 antigen Prognosis p53癌抑制遺伝子 PCR‐SSCP Ki‐67抗原 Oncogene Apotosis Lung Cancer L-myc p-53

项目摘要

In order to investingate the mechanism of apoptosis induced by p53 gene and myc genes, comparative analysis of gene abnormality in cancer patients was planed. Sesitivity test against anticaner drugs was also planed by using the cell lines which was transfected 1-myc gene.1) Mutation of P53 gene was screened by PCR-SSCP method and sequeced. : In 47 primary non-small cell lung cancer cases, 12 cases (25.5%) were mutated and 3 of them had G to T transversions which is said to be related to tobcco smoking. There was no correlation between P53 mutation and TNM factors nor histology types nor prognosis. There was no deference between the median survival time of the chemotherapy group and the control group.2) Im munohistochemical analysis of oncoprotein produced by abnormal p53 gene and Ki-67 antigen expressed in cycling cells. : In 162 non-small cell lung cancer patients surgically treated in our institute, immunohistochemical analysis using DO-7 and MIB-1 was performed with resected tumor specimens. Although no significant relation between labeling indices and pathological stage was demonstrated in both antigens, DO-7 positive tumos showed significantly lower post-operative survival rate than negative tumors in curative resection cases (p<0.05). And significant correlation between the expression of DO-7 and MIB-1 were observed (p<0.05).3) Gene transfection with lung cancer cell line. : To simplify the experiment, we are selecting cell lines which showsaberrantp53 gene without overexpression of myc gene. We planed to transefct myc gene to these cell lines and to undergo sensitivity test against etoposide with these transfected cell lines. First, We analyzed seven cell lines of human lung cancer and in five out of seven we found abnormality in p53 gene by PCR-SSCP method. Further experiment are on-going to check overexpression of myc gene with thses cell lines using northern blotting analysis.

为了探讨p53基因和myc基因诱导细胞凋亡的机制，拟对癌症患者基因异常进行比较分析。并利用转染1-myc基因的细胞系进行抗癌药物敏感性试验。1)采用PCR-SSCP方法筛选P53基因突变并测序。：在 47 例原发性非小细胞肺癌病例中，12 例（25.5%）发生突变，其中 3 例出现 G 到 T 颠换，据说与吸烟有关。 P53 突变与 TNM 因素、组织学类型和预后之间没有相关性。化疗组与对照组的中位生存时间无差异。2)循环细胞中异常表达的p53基因和Ki-67抗原产生的癌蛋白的免疫组化分析。：在本研究所接受手术治疗的 162 例非小细胞肺癌患者中，对切除的肿瘤标本进行了使用 DO-7 和 MIB-1 的免疫组织化学分析。尽管两种抗原的标记指数与病理分期之间没有显着关系，但在根治性切除病例中，DO-7阳性肿瘤的术后生存率显着低于阴性肿瘤（p<0.05）。 DO-7与MIB-1的表达存在显着相关性(p<0.05)。3)肺癌细胞系基因转染。：为了简化实验，我们选择显示 aberrantp53 基因但不过度表达 myc 基因的细胞系。我们计划将myc基因转染到这些细胞系中，并用这些转染的细胞系进行依托泊苷敏感性测试。首先，我们分析了七种人类肺癌细胞系，通过PCR-SSCP方法发现七种细胞系中的五种p53基因异常。进一步的实验正在进行中，使用 Northern blotting 分析检查这些细胞系中 myc 基因的过度表达。