| Project/Area Number |
06671328
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Thoracic surgery
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| Research Institution | Chiba University |
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Principal Investigator |
SHIBA Mitsutoshi Chiba Univ School of Medicine Assistant, 医学部, 助手 (20162620)
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| Co-Investigator(Kenkyū-buntansha) |
MITSUNAGA Shinichiro Chiba Univ School of Medicine Assistant, 付属病院, 助手 (50272331)
YAMAGUCHI Yutaka Chiba Univ School of Medicine Proffesor, 医学部, 教授 (80009448)
田宮 敬久 千葉大学, 医学部, 助手 (10251167)
川野 裕 千葉大学, 医学部, 助手 (40251160)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | 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 |
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| Research Abstract |
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.
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