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

最初，我们尝试建立多种肺癌细胞系。先前的实验表明，在胶原蛋白中混合50%的基质对细胞系的建立最有效。此外，在上述混合物中加入10%的患者血清，更容易建立细胞系。用这种方法建立了3种AAH，这是第一次在胶原凝胶中成功建立AAH细胞系，然后我们成功地培养了近10次。在这个阶段，有可能分析p53的突变。但3例均为野生型p53基因，未发现p21等p53相关基因异常。我们还从病理学的角度建立了肺癌细胞系。成功地建立了10株I期肺癌细胞株和9株II期肺癌细胞株。我们试图在手术前进行化疗的III期肺癌中建立13个细胞系，相比之下，由于大部分部位是坏死区域，仅建立了4个细胞系。然后，从转移区标本中建立了6种StageIV细胞系。从这些角度来看，我们检查p53的突变，不是从切除标本中提取的DNA，而是从这些细胞系中，这对于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。而且，无病生存和p53突变之间没有明显的关系。另一方面，在ⅱ期和ⅳ期肺癌复发患者中，我们没有发现抗癌药物敏感性与使用相同药物对患者的影响有显著关系。

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.