Analysis on rearrengement of chromosomal structure using spectral kartotyping method

光谱卡罗法分析染色体结构重排

• 批准号: 13672100
• 负责人: ITA Masamichi
• 金额: $ 2.69万
• 依托单位: Yamaguchi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672100/
• 关键词: spectral karyotyping; oral cancer cell line; squamous cell carcinoma; isochromosome; Spectral karyotyping; oral cancer; Squamous cell carcinoma; Spectral Karyotyping; SKY法; 口腔扁平上皮癌; 染色体構造異常

It has said that carcinogenesis associates with chromosomal instability. In order to detect the abnormal changes of chromosomal structure, we employed spectral karyotyping method that was recently developed. We analysed for 11 oral aquamous cell carcinoma cell lines (KM-Z, KM -3, KM-5, Sa-3, H-1, TYS, BHY, HN, OSC30, OSC70, SAS). Metaphase specimen of chromosome needs for this experiment, and we changed and mixture ratio of hypotonic solution (three parts of 0.075M potassium chloride and one pert of 0.9% civic acid). Moreover HANABI, metaphase spreader machine, provided proper dispersion of chromosemes. Numerical aberration of chromosome number and changes of chromosomal structure were analyzed with SKY View Computer software. DNA index was measured, by flowcytometry to compare with chromosome numberThe number of chromosome showed dispersion ; KM-2 cells showed the smallest number of chromosome (49-53 chromosomes), on the other hand, Sa-3 cells revealed the largest number of it (98-100 chromosomes). DNA index ranged from 1.35 to 1.64. Although many cell lines showed the relation between chromosome number and DNA index, a few cell lines such as Sa-3 did not show ; DNA index of Sa-3 containing 98-100 chromosomes was 1.48. A number of structural changes of chromosomes like deletion ranged from 16 to 33. These results suggested that carcinogenesis seems to relate to chromosomal structural rearrangement and abnormal segregation. Nine cell lines disclosed isochromosome of short arm of chromosome 5. It was confirmed that there is i(5)(p10) at 5p15 region with FISH using BAC clone. We supposed that this region relates to increase of DNA copy number

据说致癌与染色体不稳定有关。为了检测染色体结构的异常变化，我们采用了最近发展起来的光谱核型分析方法。我们分析了 11 种口腔水生细胞癌细胞系（KM-Z、KM -3、KM-5、Sa-3、H-1、TYS、BHY、HN、OSC30、OSC70、SAS）。本实验需要染色体中期标本，我们改变了低渗溶液（三份0.075M氯化钾和一份0.9%柠檬酸）的混合比例。此外，HANABI，中期分散机，提供了染色体的适当分散。利用SKY View Computer软件分析染色体数目的数值畸变和染色体结构的变化。测定DNA指数，通过流式细胞仪与染​​色体数目进行比较，染色体数目呈现分散性； KM-2细胞显示的染色体数量最少（49-53条染色体），而Sa-3细胞显示的染色体数量最多（98-100条染色体）。 DNA指数范围为1.35至1.64。虽然许多细胞系显示出染色体数目和DNA指数之间的关系，但少数细胞系如Sa-3没有显示；含有98-100条染色体的Sa-3的DNA指数为1.48。染色体的一些结构变化，如缺失，范围从16到33条不等。这些结果表明，致癌作用似乎与染色体结构重排和异常分离有关。 9个细胞系显示5号染色体短臂等染色体。使用BAC克隆进行FISH证实5p15区域存在i(5)(p10)。我们推测这个区域与DNA拷贝数的增加有关

项目成果

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Wojciech Gorezyca：“一种新型抗癌核糖核酸内切酶 ONCONASE 与分化剂协同诱导细胞凋亡（综述）”细胞计数法。

Yukou Saito: "A signal pathway by a new synthetic lipid A analog, ONO-4007,on RAW264.7 cells"Anti-cancer Drug. 13. 1069-1075 (2002)

Yukou Saito：“新型合成脂质 A 类似物 ONO-4007 在 RAW264.7 细胞上的信号通路”抗癌药物。

Masamichi Ita: "A case of maxillary sinus plasmacytoma and analysis cellular biological features"The bulletin of the yamaguchi medical school. 49. 107-113 (2002)

伊田正通：《上颌窦浆细胞瘤一例及细胞生物学特征分析》山口医学院通报。

Masamichi Ita: "A case of maxillary sinus plasmacytoma and analysis of cellular biological features"The bulletin of the yamaguchi medical school. 49. 107-113 (2002)

伊田正通：《上颌窦浆细胞瘤一例及细胞生物学特征分析》山口医学院通报。

Aoe K: "Gingival metastasis as initial presentation of small cell carcinoma pf the lung"Anticancer Research. 23. 4187-4190 (2003)

Aoe K：“牙龈转移是小细胞肺癌的最初表现”抗癌研究。