YAMANDA Takeshi Shinshu University School of Medicine, Assistant Professor, 医学部・第二外科, 助手 (20273089)
FUJIMORI Minoru Shinshu University School of Medicine, Assistant Professor, 医学部・附属病院・第二外科, 助手 (00262725)
FUKUSHIMA Yoshimitsu Shinshu University School of Medicine, Professor, 医学部・衛生学, 教授 (70273084)
小出 直彦 信州大学, 医学部・附属病院, 助手 (10283269)
|Budget Amount *help
¥12,800,000 (Direct Cost : ¥12,800,000)
Fiscal Year 1999 : ¥1,400,000 (Direct Cost : ¥1,400,000)
Fiscal Year 1998 : ¥1,200,000 (Direct Cost : ¥1,200,000)
Fiscal Year 1997 : ¥10,200,000 (Direct Cost : ¥10,200,000)
We studied if it is possible to detect an identical mutation in free plasma DNA in the peripheral blood and lung cancer tissue. Fifty seven patients were Studied, including 34 adenocarcinomas and 23 squamous cell carcinomas of the lung. Tissue samples from tumor and corresponding normal lesions were sampled. Pre-operative blood samples were collected for free plasma DNA. Using 4 kinds of micro-satellite DNA marker(D3S1284, D3S1300, TP53, D21S1245), we found 53%, 18%, 29%, 0% mutations respectively in adenocarcinomas, and 9%, 4%, 6%, 0% mutations respectively in squamous cell carcinomas. Although we investigated plasma DNA in order to find identical mutation that is found in tumor tissue, we could not detect satisfactory results. It is supposed that micro-satellite DNA analysis in plasma DNA may not be suitable for detecting mutations in non-small cell lung cancer.
We also investigated p53 point mutations in free plasma DNA in lung cancer patients. We screened p53, from exon 5 through 8, in 12 non-small-cell lung cancer tissue by SSCP analysis. Eight cases out of 12, we found mutations in tumor tissue, but we could not get appropriate PCR products from plasma DNA for further analyses.
Comparative genomic hybridization (CGH) method was also carried out to find out new mutations in lung cancer. We found few clear results of hybridization, but in most of the patients, we could not get clear, repetitive, and stable results, so that further analyses other than our method should be done.
Through these studies supported by this grant, our technique and equipment on molecular research were quiet valuable for finding new knowledge in other diseases besides lung cancer, such as gastric cancer, breast cancer, and cardiac myxoma.