JaLCDOI 10.18926/AMO/30772
フルテキストURL fulltext.pdf
著者 Pu, Hong| Tsuji, Toshiya| Kondo, Asami| Fushimi, Kazuo| Ohashi, Ryuichiro| Inoue, Yusuke| Mimura, Tetsushige| Hamazaki, Keisuke| Miyazaki, Masahiro| Namba, Masayoshi|
抄録 <p>Characteristics of human hepatoma cell lines with the wild-type p53 were compared with those of human hepatoma cell lines with the mutant-type p53. The p21 protein located downstream of p53 was expressed in cell lines with the wild-type p53 but was not expressed in cell lines with the mutant-type p53. As to other tumor suppressor genes such as p16 and p27, there was no difference in their expression between both types of cell lines. In addition, no marked difference was observed in the activities of CDK2 and CDK4 between cell lines with the wild-type and the mutant-type p53. Phosphorylated Rb protein was detected in all cell lines except the HLE line, indicating that this cell line may have a deletion of and/or a mutation of the Rb gene. These results indicate that abnormalities of tumor suppressor genes other than p53, p16, p27, and Rb may be involved in hepatocarcinogenesis. The population doubling time of the wild-type p53 cells was significantly longer than that of the mutant p53 cells. Neither type of cell line showed a specific chromosome distribution which would indicate karyotype instability. The cell lines expressing the wild-type p53 produced tumors at lower frequency than those with the mutant p53 gene. Although there was no significant difference in effects of TGF-&#946;1, EGF, cholera toxin, and db-cAMP on cell growth between the two types of cells, all three cell lines with the wild-type p53 were resistant to cytotoxicity of TNF-&#945;, while two of the three with the mutant p53 were very sensitive to its cytotoxic effects.</p>
キーワード hepatoma p53 p21?p16?p27?Rb TNF-?
Amo Type Article
発行日 1997-12
出版物タイトル Acta Medica Okayama
51巻
6号
出版者 Okayama University Medical School
開始ページ 313
終了ページ 319
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 9439773
Web of Sience KeyUT 000071183400004
JaLCDOI 10.18926/AMO/30502
フルテキストURL fulltext.pdf
著者 Kosaka, Tsunenori| Fukaya, Ken-ichi| Tsuboi, So| Pu, Hong| Ohno, Tadao| Tsuji, Takao| Namba, Masayoshi|
抄録 <p>The sensitivity of five kinds of cytotoxicity assays using ethanol on human hepatoblastoma cells (HUH-6 line), which were cultured as monolayers or spheroids, was compared. Ethanol was chosen as a test because it acts on cell membranes directly without being metabolized and exerts its cytotoxicity. The assay methods used were as follows: 3- (4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH), colony formation, cell growth and DNA assays. The sensitivity of the assays was: LDH &#60; DNA &#60; cell growth &#60; MTT &#60; colony formation. LDH assay had the advantage that the same culture could be used for multiple assays, but when a small number of cells were assayed, no significant increase in the release of LDH was detected in the assay cultures compared with the control cultures. Although the DNA and cell growth assays were more sensitive than the LDH assay, the extent of cell damage may be underestimated because the damaged cells and DNA present in the cultures are included in the assay samples. On the other hand, both MTT and colony formation assays showed a high sensitivity. The MTT assay was done within 24 h after ethanol was added to the cultures and was applicable to both monolayer and spheroid cultures, while the colony formation assay required 1-2 weeks and it was applicable only to monolayer cultures. Taken together, the MTT assay was the most suitable method to evaluate the cytotoxic effects of ethanol on HUH-6 cells cultured as either monolayers or spheroids.</p>
キーワード human hepatoblastoma monolayer culture spheroid culture cytotoxicity
Amo Type Article
発行日 1996-06
出版物タイトル Acta Medica Okayama
50巻
3号
出版者 Okayama University Medical School
開始ページ 151
終了ページ 156
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 8805855
Web of Sience KeyUT A1996UU60400006
JaLCDOI 10.18926/AMO/30490
フルテキストURL fulltext.pdf
著者 Kosaka, Tsunenori| Tsuboi, So| Fukaya, Ken-ichi| Pu, Hong| Ohno, Tadao| Tsuji, Takao| Miyazaki, Masahiro| Namba, Masayoshi|
抄録 <p>&#60;P&#62;Spheroid cultures of human hepatoblastoma cells (HuH-6 line) were established by rotating 3 x 10(6) cells/3 ml culture medium in 25-ml Erlenmeyer flasks on a gyratory shaker. The size of the spheroids rapidly increased until 4 days of culture, and thereafter their size gradually increased until 8 days of culture. A considerable amount of lactate dehydrogenase (LDH) was detected in the culture medium at 24h after seeding because of cell damage by subculturing, but thereafter the amount released was small, indicating that the spheroids were in healthy condition. Albumin production, one of the differentiated functions of hepatocytes, was higher in spheroid cultures than in monolayer cultures. Using this spheroid culture model, the cytotoxic effects of alcohols on HuH-6 cells were studied by measuring the activity of LDH released in the medium from damaged cells. The results indicate that the increasing order of toxicity of the alcohols was as follows: methanol &#60; ethanol &#60; propanol.&#60;/P&#62;</p>
キーワード human hepatoblastoma spheroid cultures albumin alcohols LDH
Amo Type Article
発行日 1996-04
出版物タイトル Acta Medica Okayama
50巻
2号
出版者 Okayama University Medical School
開始ページ 61
終了ページ 66
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 8744930
Web of Sience KeyUT A1996UJ08100001