1990 Fiscal Year Final Research Report Summary
Cell Culture and Molecular Biologicals Study for Human Esophageal Carcinom
| Project/Area Number |
63570639
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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 |
Digestive surgery
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| Research Institution | Yamaguchi University |
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Principal Investigator |
MURAKAMI Takuo Yamaguchi University School of Medicine. Second Department of Surgery. Associate Professor, 医学部, 助教授 (10091216)
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| Co-Investigator(Kenkyū-buntansha) |
JOHONO Norifumi Yamaguchi University School of Medicine. Hospita Second department of Surgery., 医学部附属病院, 助手 (70196666)
OKA Masaaki Yamaguchi University School of Medicine. Second department of Surgery., 医学部, 講師 (70144946)
SHIMIZU Ryouichi Yamaguchi University School of Medicine. Hospital. Second department of Surgery., 医学部附属病院, 助手 (30187471)
KUSANAGI Hiroshi Yamaguchi University School of Medicine. Hospital Second department of Surgery.
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| Project Period (FY) |
1988 – 1990
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| Keywords | Esophageal cancer / cell line / Southern blotting / erb-B / myc / EGF / serum-free culture / Etoposide |
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| Research Abstract |
Six squamous cell carcinoma cell lines, YES-1, YES-2, YES-3, YES-4, YES-5 and YES-6, were established from human esophageal carcinoma.
YES-1 and YES-2 were established from subcutaneus small tumor implanted into nude mice, which had been transplanted from a surgical specimen obtained from esophageal cancer patients.
YES-3 was established from pleural effusion of the carcinomatous pleuritis for carcinoma of the esophagus. The remaining YES-4, YES-5 and YES-6 were derived from a surgical specimen.
Deoxyribonucleic acids were extracted from these 6 human esophageal carcinoma cell lines using the phenol-chloroform method. The C-erb B gene and the C-myc gene were examined for possible structural changes in them by Southern blot hybridization. The C-erb B gene was amplificated about ten folds in YES-1 and amplification of C-myc could not be detected in these six cell lines.
In our laboratory, we have a problem concerning the manipulation of Southern blot hybridization, and we must check the resu
… More
lt again.
The amplification of various oncogenes will be examined by using Northern blotting method, when the manipulation procedure has been mastered.
We are examining the role of EGF in vitro and in vivo using YES-2, which had the shortest dubbing time among the six cell lines. We are going to examine it using five other cell lines in near future. Additionally we will determine the binding of EGF to membrane fractions.
YES-1 and YES-2 were adapted to the serum-free medium. These two lines may produce some substances, such as growth factors.
We are now examining four other cell lines which have been adapted to the serum-free medium.
The influence on cell proliferation of etoposide which is an anticancer drug was examined in YES-1 cells using flow cytometric analysis.
Furthermore, we have scheduled to examine various kinds anticancer drugs, such as cisdiamine dichloroplatinum, vindesin, adriamycin and 5-fluorouracil sensitivity by using these 6 cell lines.
These 6 cell lines may provide useful models for the study of human esophageal carcinoma. Less
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