Study on Oncogene in Juvenile Hepatocellular Carcinoma in China
Grant-in-Aid for Overseas Scientific Survey.
|Section||Special Cancer Research|
|Research Institution||Kyushu University|
NAKAYAMA Fumio Kyushu University Faculty of Medicine, Professor, 医学部, 教授 (70038652)
夏 亮芳 貴陽医科大学, 教授
郭 仁宣 中国医科大学, 副教授
張 振〓 協和医科大学, 副教授
姚 来礼 新彊医学院, 教授
曹 綉虎 中山医科大学, 教授
朱 予 協和医科大学, 教授
沈 魁 中国医科大学, 教授
関口 睦夫 九州大学, 医学部, 教授 (00037342)
XIA Liang Fang Guizhou Medical College, Professor
GUO Ren Xuan China Medical College, Associate Professor
ZAHNG Zhen Xuan Peking Union Medical College, Associate Professor
YAO Ping Li Xinjiang Medical College, Professor
ZHU Yu Peking Union Medical College, Professor
TSAO Siu Hu Sun Yat Sen Univ. Med. Sciences, Professor
SEKIGUCHI Mutsuo Kyushu University Faculty of Medicine, Professor
SHEN Kui China Medical College, Professor
|Project Fiscal Year
1989 – 1990
Completed(Fiscal Year 1990)
|Budget Amount *help
¥7,000,000 (Direct Cost : ¥7,000,000)
Fiscal Year 1990 : ¥4,000,000 (Direct Cost : ¥4,000,000)
Fiscal Year 1989 : ¥3,000,000 (Direct Cost : ¥3,000,000)
|Keywords||Hepatocellular carcinoma / China / Epidemiology / Oncogene / DNA / Primary transformant / Secondary transformant / 肝細胞癌 / 中国 / 疫学 / 癌遺伝子 / 一次トランスフォ-マント / 二次トランスフォ-マント|
In order to identify oncogenes in hepatocellular carcinoma in China, several institutes in China have been visited including China Medical College, Guiyang Medical College and Xinjiang Medical College. Patient's record, resected liver specimen and histology have been examined and documented. Total of 17 cases have been collected avoiding those in which transarterial embolisation and/or chemotherapy have been used - the former causes necrosis of tumor mass and the latter possible derangement to the DNA.
Frozen resected liver specimen brought back to Kyushu University Faculty of Medicine, Department of Surgery I have been used for identification of oncogenes by isolating DNA.
From 14 specimens, high molecular weight DNAs were prepared. These cellular DNA samples (30 mug/dish) were used to transfect mouse NIH/3T3 cells together with pSV2hph (1mug/dish) using modified calcium phosphate precipitation method. The cells were incubated for 13 hours with the DNA precipitate and then freed from th
e DNA. Thirty six hours after transfection, they were divided into three dishes, and refed with Dulbecco's modified Eagle's medium supplemented with 5% calf serum and Hygromycin B (150mug/ml) every 4 days. Morphologically transformed foci that appeared after incubation 14-21 days in 8 initial DNA samples (primary foci) were scored and isolated. The DNAs of the primary foci were analyzed by Southern blot hybridization with human placenta DNA as a probe for the presence of human-specific repeated sequences (Alu seq.). Fourteen DNA samples from primay foci originating from 7 initial DNA preparations retained large amounts of the human DNA sequences (primary transformants).
The DNAs of the primary transformants were then used for the next round of transfection to obtain secondary transformants. Twelve secondary transformants were obtained independently from 9 different primary transformants, which originated from 3 different samples. The results of Southern blot hybridization of the secondary transformants gave patterns different from those of K-ras and H-ras. These secondary transformants did not give hybridization signals when N-ras, Ica and hst-1 DNAs were used as probes. Thus there might be unknown oncogenes in these secondary transformants. Presently we are screening of the lambda phage and the cosmid libraries prepared from the secondary transformants.
Research Output (4results)