Effect of 3′-Methyl-4-dimethylaminoazobenzene on liver cells from adult rat in primary culture

Miyazaki, Mikihiro; Wahid, Syarifuddin; Matsuura, Koji; Tsunashima, Masakazu; Nakabayashi, Hidekazu; Miyano, Keiko; Sato, Jiro

doi:10.1007/bf01850987

Cited by 7 publications

(3 citation statements)

References 9 publications

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“…Since it has been reported that the gamma-GTP activity per cell was independent of cell density (Sakakibara and Tsukada, 1980), the assay for gamma-GTP was carried out on the cultures in confluent state. As reported previously (Miyazaki et al, 1982), the cytochemical assay for gamma-GTP on the cultures fixed with cold acetone was carried out according to the method of Albert et al (1961) using N-gamma-L-glutamyl-alphanaphthylamide (Sigma Chemical Co., St. Louis, MO, USA) as a substrate, fast garnet GBC salt (Sigma Chemical Co., St. Louis, MO, USA) as a diazonium coupling reagent and glycylglycine (Sigma Chemical Co., St. Louis, MO, USA) as an acceptor for acceleration of the enzymatic reaction. Nuclei of cultured cells were stained with hematoxylin.…”

Section: Cytochemical Assay For Gamma-gtpsupporting

confidence: 64%

Expression of gamma‐glutamyltranspeptidase in cultures of spontaneously and chemically transformed rat liver cells

Miyazaki

Wahid

Miyano

et al. 1983

Intl Journal of Cancer

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The expression of gamma-glutamyltranspeptidase (gamma-GTP) and the relationship between this enzyme activity and tumorigenicity were studied in 38 tumorigenic and non-tumorigenic epithelial cell lines derived from livers of normal and aminoazo-dye-fed rats. Nine of 22 non-tumorigenic epithelial cell lines exhibited gamma-GTP-positive cells, whereas 9 of 16 tumorigenic epithelial cell lines did not exhibit gamma-GTP-positive cells. This finding revealed that no correlation existed between the acquisition of gamma-GTP activity and tumorigenicity. None of 10 spontaneously transformed epithelial cell lines, which consisted of 8 tumorigenic and 2 non-tumorigenic cell lines, exhibited gamma-GTP-positive cells. On the other hand, 16 of 28 transformed epithelial cell lines, which were derived from aminoazo-dye-fed rat livers and aminoazo-dye-treated cultures of normal rat liver cells, included gamma-GTP-positive cells in various percentages. As described above, in contrast to in vitro spontaneous transformation of rat liver cells, it is obvious that the carcinogen-induced transformation of rat liver cells in vitro as well as in vivo was frequently accompanied by the acquisition of gamma-GTP-activity. Therefore, it is concluded that the expression of gamma-GTP in liver cells may be mainly associated with exposure to chemical carcinogens.

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Section: Cytochemical Assay For Gamma-gtpsupporting

confidence: 64%

Expression of gamma‐glutamyltranspeptidase in cultures of spontaneously and chemically transformed rat liver cells

Miyazaki

Wahid

Miyano

et al. 1983

Intl Journal of Cancer

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“…As reported previously, liver cells were isolated from adult rats by trypsin and collagenase perfusion techniques [15,17]. The perfusate used in this study were as follows.…”

Section: Preparation and Primary Culture Of Isolated Liver Cellsmentioning

confidence: 99%

Comparison of cytologic and biochemical properties between liver cells isolated from adult rats by trypsin perfusion and those isolated by collagenase perfusion

et al. 1984

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Primary cultures of liver cells isolated from adult rats by trypsin and collagenase perfusion techniques were carried out to compare cytologic and biochemical properties between the differently prepared cells. Trypsin-dispersed cells consisted of comparatively smaller cells, whereas collagenase-dispersed cells consisted of larger cells. The cell attachment efficiency on culture day 1 was about twice as high in the liver cells prepared with collagenase than those prepared with trypsin. Mature hepatocytes isolated by collagenase perfusion could be maintained in the primary culture for a longer period than those isolated by trypsin perfusion. Epithelial-like clear cells started to grow much earlier in the primary culture of the trypsin-dispersed liver cells than in that of the collagenase-dispersed liver cells. Earlier proliferation of epithelial-like clear cells could not be induced by in vitro trypsinization of the collagenase-dispersed liver cells. Both kinds of enzymatically prepared liver cells showed albumin production and exhibited glucose 6-phosphatase (D-glucose-6-phosphate phosphohydrolase, EC 3.1.3.9, G6Pase) and tyrosine aminotransferase (L-tyrosine: 2-oxoglutarate amino-transferase, EC 2.6.1.5, TAT) activities for 1 week in the primary culture. Albumin production was higher in the liver cells prepared with collagenase than those prepared with trypsin, whereas G6Pase activity was almost the same between them. TAT activity up to culture day 2 was about 3-fold higher in the liver cells prepared with collagenase than in those prepared with trypsin. Combined supplementation of dexamethasone (1 X 10(-5)M) and insulin (10 micrograms/ml) consistently improved the cell attachment efficiency and was very effective in the maintenance of mature hepatocytes in both types. Furthermore, these hormones enhanced the albumin production and TAT activity in both types.

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“…Similar doses of AO were found to be cytotoxic in Sertoli germ cells (Mishra et al 2009); and for BY in primary hepatocytes (Miyazaki et al 1982). Significant loss of membrane integrity and leakage of LDH in the medium was observed by AO and BY in a dose-and time-dependent manner.…”

Section: Discussionmentioning

confidence: 90%

Role of ErbB2 mediated AKT and MAPK pathway in gall bladder cell proliferation induced by argemone oil and butter yellow

et al. 2012

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The effect of noncytotoxic doses of argemone oil (AO) and butter yellow (BY), the common adulterants in edible oil, on free radical generation and signaling pathway for cell proliferation in primary cells of gall bladder (GB) was undertaken. AO and BY showed no cytotoxicity at 0.1 μl/ml and 0.1 μg/ml concentration, respectively. AO caused significant increase in ROS after 30 min and RNS after 24 h in GB cells while no change was observed following BY treatment. Enhanced level of COX-2 was observed following AO (0.1 μl/ml) and BY (0.1 μg/ml) treatment to cells for 24 h. AO treatment caused phosphorylation of ErbB2, AKT, ERK, and JNK along with increased thymidine uptake indicating cell proliferation ability in GB cells. BY treatment also showed significant expression of these proteins with the exception of phosphorylated JNK. These results suggest that AO and BY have cell proliferative potential in GB cells following up-regulation of COX-2 and ErbB2; however, their downstream signaling molecules and free radical generation have differential response, indicating that the mechanism of proliferation is different for both compounds and may have relevance in gall bladder cancer.

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Effect of 3′-Methyl-4-dimethylaminoazobenzene on liver cells from adult rat in primary culture

Cited by 7 publications

References 9 publications

Expression of gamma‐glutamyltranspeptidase in cultures of spontaneously and chemically transformed rat liver cells

Expression of gamma‐glutamyltranspeptidase in cultures of spontaneously and chemically transformed rat liver cells

Comparison of cytologic and biochemical properties between liver cells isolated from adult rats by trypsin perfusion and those isolated by collagenase perfusion

Role of ErbB2 mediated AKT and MAPK pathway in gall bladder cell proliferation induced by argemone oil and butter yellow

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