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Development of CD-DST sensitivity test and analysis of relationship between p53 related genes and prognosis.

CD-DST敏感性试验的开展及p53相关基因与预后关系分析。

基本信息

批准号：
14571168
负责人：
ISHIDA Hisao
金额：
$ 2.24万
依托单位：
The Tazuke Kofukai
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571168/
关键词：
anti cancer drug CD-DST matrigel p53

项目摘要

Initially, we tried to establish many kinds of cell lines of lung cancer. The previous experiments showed that mixing 50% matrigel in this collagen was the most efficient for establishment of the cell lines. Furthermore, adding 10% of patients' serum in the above mixture made it easier to establish the cell line. Using this method, three kinds of AAH were established, which was the first success of establishment AAH cell lines in the collagen gel, and then we succeeded in almost 10 times cultures. At this stage, it was possible to analyze the mutation of p53. But, all three cases had wild type of p53 gene, and abnormality of p53 related genes for example, p21was not found. We also established cell lines on the lung cancers from the point of view of pathology. It was succeeded to establish 10 cell lines in Stage I lung cancer and 9 cell lines in Stage II. We tried to establishl3 cell lines in Stage III lung cancer that did chemotherapy before the operation, in contrast, only 4 cell lines were established because most of the parts were necrotic region. Then, 6 kinds of cell lines in StageIV were established from the specimens of metastasis region. From these points, we examined mutations of p53, not from extracted DNA from the resected specimens, but from those cell lines, which is important to avoid the contamination of normal cells. However, we could not find any relationships between mutations of p53 and stages. And also, no significant relationships between diseases free survival and p53 mutations were found. On the other hand, we could not find the significant relationship of anticancer drug sensitivity and the effect on the patients using the same drugs in the patients with the relapse of StageIII and StageIV lung cancer.

最初，我们试图建立多种肺癌细胞系。以往的实验表明，在该胶原中混合50%的基质蛋白是建立细胞系的最有效的方法。此外，在上述混合物中加入10%的患者血清可以更容易地建立细胞系。用这种方法建立了3种AAH，这是首次在胶原胶中成功建立AAH细胞系，随后又成功地进行了近10次培养。在这个阶段，分析p53的突变是可能的。但3例均为野生型p53基因，未发现p21等p53相关基因异常。我们还从病理学角度建立了肺癌细胞系。成功地建立了10个I期肺癌细胞系和9个II期肺癌细胞系。我们尝试建立了3个术前化疗的III期肺癌细胞系，而由于大部分部位是坏死区，所以只建立了4个细胞系。然后，从转移灶标本中建立了6种IV期细胞系。从这些方面，我们检查了p53的突变，不是从切除的标本中提取的DNA，而是从这些细胞系中提取的DNA，这对避免正常细胞的污染很重要。然而，我们没有发现P53突变与分期之间的任何关系。此外，无病生存期与P53基因突变之间也没有发现明显的关系。而在复发的III期和IV期肺癌患者中，未发现抗癌药物敏感性与使用相同药物的患者的疗效之间的显著关系。