Different Expression of Esterase Variants in Rat Hepatic and Hepatom a-Derived Cell Lines Detected by Electrophoresis

1995 ◽  
Vol 50 (9-10) ◽  
pp. 664-668 ◽  
Author(s):  
Adel S. Afify ◽  
Yoshimitsu Yamazaki ◽  
Yu-ichi Kageyama ◽  
Shiro Yusa ◽  
Yoshikatsu Ogawa ◽  
...  
Keyword(s):  
Cell Lines ◽  
Rat Liver ◽  
Hepatoma Cell ◽  
Polyacrylamide Gel Electrophoresis ◽  
Liver Homogenate ◽  
Normal Liver ◽  
Liver Parenchyma ◽  
Hepatoma Cell Lines ◽  
Normal Rat ◽  
Transformed Cell Lines

Abstract Esterases in nine rat hepatic and hepatoma-derived cell lines and normal rat liver homogenate were detected by polyacrylamide gel electrophoresis coupled with active staining with a-naphthyl acetate or butyrate as a substrate. The esterase band patterns of the non-cancerous and oncogene-transformed cell lines were alike, but different from those of hepatoma cell lines and normal rat liver homogenate. The former groups of cells might have completely lost the characteristics of rat liver parenchymal cells, or else they might have their origin at cells other than liver parenchyma. The esterase patterns of the hepatoma cell lines (e.g., McA-RH7777) and the normal rat liver highly resembled with each other, exemplifying the slight biochemical deviation of cancer from normal cells. However, two-dimensional electrophoretogram for the McA-RH7777 cell line showed a prominent esterase spot {p/ 6.0-Mr 110 kDa) that was lacking in the normal liver. This result indicates that there is invariably some change in esterase expression between the cancer cells and the normal liver cells

Cell surface topography in adhesion and detachment

1978 ◽  
Vol 36 (2) ◽  
pp. 300-301
Author(s):  
T.D. Allen
Keyword(s):  
Cell Lines ◽  
Thin Sheet ◽  
Liver Parenchyma ◽  
Soft Agar ◽  
Transformed Cells ◽  
Normal Cells ◽  
Epithelial Cell Lines ◽  
Normal Rat ◽  
Transformed Cell Lines ◽  
Peripheral Cytoplasm

The effects of trypsin and EGTA were investigated in parallel on two epithelial cell lines: a normal rat-liver parenchyma line (Fig. 1) and a spontaneously transformed line which had been subcloned in soft agar. The basic ultrastructural differences between these two cell lines has already been reported; briefly, the transformed cells, although typically epithelial in morphology, lack the well ordered microtubular and microfilamentous organisation of the normal cells but exhibit a marked increase in the number of surface microvilli.The reaction of each cell line to trypsination was broadly similar between the normal and transformed cell lines, with the transformed cells rounding up and detaching at a slightly quicker rate than the normals. In both cases the central region of the cell became domed with the peripheral cytoplasm being withdrawn in a thin sheet, some of which remained extended in short retraction fibrils (Fig. 2). Collection of the detached cells (after fixation in suspension) by aspiration onto silver filters showed the increased density of microvilli on the transformed cells to be maintained.

Molecular cloning of cDNA for rat argininosuccinate lyase and its expression in rat hepatoma cell lines

1986 ◽  
Vol 6 (5) ◽  
pp. 1722-1728
Author(s):  
M A Lambert ◽  
L R Simard ◽  
P N Ray ◽  
R R McInnes
Keyword(s):  
Cell Lines ◽  
Rat Liver ◽  
Transcriptional Activity ◽  
Hepatoma Cell ◽  
Cdna Clones ◽  
Argininosuccinate Lyase ◽  
Hepatoma Cell Lines ◽  
Liver Cdna Library ◽  
Nuclear Processing ◽  
Rat Hepatoma

Using antibody and plaque hybridization screening, we isolated rat argininosuccinate lyase (AS lyase) cDNA clones from a liver cDNA library prepared in the phage expression vector lambda gt11. Five overlapping cDNAs covering 1.7 kilobases of the estimated 2.0-kilobase AS lyase mRNA were characterized and confirmed as AS lyase sequences by hybrid selection. We examined the differential expression of AS lyase in rat liver and four rat hepatoma cell lines (7800C1, H4, HTC, and MH1C1). These cells exhibited a 60-fold range of AS lyase enzyme activity, with a direct correlation between activity, amount of AS lyase immunoreactive protein, and quantity of specific AS lyase mRNA. These observations suggest that the differences in AS lyase expression between rat liver and the hepatoma cell lines result from variations in AS lyase transcriptional activity or alterations in nuclear processing of AS lyase RNA.

scholarly journals Heparin and Liver Heparan Sulfate Can Rescue Hepatoma Cells from Topotecan Action

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
József Dudás ◽  
József Bocsi ◽  
Alexandra Fullár ◽  
Kornélia Baghy ◽  
Tibor Füle ◽  
...  
Keyword(s):  
Liver Cancer ◽  
Heparan Sulfate ◽  
Cell Lines ◽  
Topoisomerase I ◽  
Hepatoma Cell ◽  
Normal Liver ◽  
Hepatoma Cell Lines ◽  
Inhibitory Compound ◽  
Hep3b Cells

Topotecan (TpT) is a major inhibitory compound of topoisomerase (topo) I that plays important roles in gene transcription and cell division. We have previously reported that heparin and heparan sulfate (HS) might be transported to the cell nucleus and they can interact with topoisomerase I. We hypothesized that heparin and HS might interfere with the action of TpT. To test this hypothesis we isolated topoisomerase I containing cell nuclear protein fractions from normal liver, liver cancer tissues, and hepatoma cell lines. The enzymatic activity of these extracts was measured in the presence of heparin, liver HS, and liver cancer HS. In addition, topo I activity, cell viability, and apoptosis of HepG2 and Hep3B cells were investigated after heparin and TpT treatments. Liver cancer HS inhibited topo I activity in vitro. Heparin treatment abrogated topo I enzyme activity in Hep3B cells, but not in HepG2 cells, where the basal activity was higher. Heparin protected the two hepatoma cell lines from TpT actions and decreased the rate of TpT induced S phase block and cell death. These results suggest that heparin and HS might interfere with the function of TpT in liver and liver cancer.

Use of Cultured Hepatoma Cell Lines in the Assessment of Aldehyde Metabolism

1992 ◽  
Vol 20 (1) ◽  
pp. 77-83
Author(s):  
Margherita Ferro ◽  
Anna Maria Bassi ◽  
Susanna Penco ◽  
Sandra Piana ◽  
Giambattista Ravera ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Hepatoma Cell ◽  
Rat Hepatocytes ◽  
Experimental Conditions ◽  
Major Pathway ◽  
Hepatoma Cell Lines ◽  
Normal Rat ◽  
Htc Cells

Aldehyde dehydrogenases (ALDH) represent a major pathway for the enzymatic removal of many potentially toxic aldehydes. The purpose of this study was to examine the basal levels of ALDH in five hepatoma cell lines chosen as representatives of three different species (man, rat, mouse) and their inducibility by some xenobiotics. Human HepG2, rat MH1C1, HTC, H4IIEC3 and mouse Hepa 1c1c7 cell lines were grown as monolayers. The ALDH activities were determined in cell homogenates from both unexposed control cultures and cells exposed to phenobarbital (PB), 3-methylcholanthrene (MC) and β-naphthoflavone (BNF). The ALDH activity was detected using benzaldehyde (BA) and propionaldehyde (PA) as substrates and both NAD and NADP as co-enzymes. Great variability in basal ALDH levels was found in the five cell lines: BA/NAD and BA/NADP enzyme activities were very high in the HTC cell line, intermediate in MH1C1 cells (near to normal rat hepatocytes) and very low in the remaining three cell lines. In HTC cells only, the PA/NAD activity was slightly induced by PB, but it remained unchanged under all the other experimental conditions. MH1C1 cells showed highly significant increases of all the activities with MC and BNF (up to 10-fold). The low basal activity of the H4IIEC3 cell line was significantly increased by MC and BNF, but only with BA/NADP. The Hepa 1c1c7 cell line responded only to BNF, as inducing compound, whereas the low basal enzyme levels of the human-derived HepG2 cell line were not significantly increased. These results suggest various applications of hepatoma cell cultures in in vitro toxicology.

scholarly journals Molecular cloning of cDNA for rat argininosuccinate lyase and its expression in rat hepatoma cell lines.

1986 ◽  
Vol 6 (5) ◽  
pp. 1722-1728 ◽  
Author(s):  
M A Lambert ◽  
L R Simard ◽  
P N Ray ◽  
R R McInnes
Keyword(s):  
Cell Lines ◽  
Rat Liver ◽  
Transcriptional Activity ◽  
Hepatoma Cell ◽  
Cdna Clones ◽  
Argininosuccinate Lyase ◽  
Hepatoma Cell Lines ◽  
Liver Cdna Library ◽  
Nuclear Processing ◽  
Rat Hepatoma

Using antibody and plaque hybridization screening, we isolated rat argininosuccinate lyase (AS lyase) cDNA clones from a liver cDNA library prepared in the phage expression vector lambda gt11. Five overlapping cDNAs covering 1.7 kilobases of the estimated 2.0-kilobase AS lyase mRNA were characterized and confirmed as AS lyase sequences by hybrid selection. We examined the differential expression of AS lyase in rat liver and four rat hepatoma cell lines (7800C1, H4, HTC, and MH1C1). These cells exhibited a 60-fold range of AS lyase enzyme activity, with a direct correlation between activity, amount of AS lyase immunoreactive protein, and quantity of specific AS lyase mRNA. These observations suggest that the differences in AS lyase expression between rat liver and the hepatoma cell lines result from variations in AS lyase transcriptional activity or alterations in nuclear processing of AS lyase RNA.

scholarly journals Basal promoter of the rat connexin 32 gene: identification and characterization of an essential element and its DNA-binding protein.

1995 ◽  
Vol 15 (3) ◽  
pp. 1439-1445 ◽  
Author(s):  
S Bai ◽  
A Schoenfeld ◽  
A Pietrangelo ◽  
R D Burk
Keyword(s):  
Cell Lines ◽  
Rat Liver ◽  
Liver Tissue ◽  
Nuclear Protein ◽  
Hepatoma Cell ◽  
Essential Element ◽  
Connexin 32 ◽  
Mobility Shift ◽  
Preferential Expression ◽  
Hepatoma Cell Lines

The connexin 32 (Cx32) gene, a member of a multigene family, is expressed preferentially in the liver. The basal promoter complex of the rat Cx32 gene was previously localized to a 146-bp region (map positions [mp] -179 to -34) immediately upstream of the first exon. To investigate the biochemical factors contributing to the basal promoter activity, nuclear protein-DNA complexes within this region (mp -177 to -106) were investigated by using a DNA mobility shift assay. Three DNA-protein binding activities, termed Cx32-B1, Cx32-B2, and Cx32-B3, were identified with nuclear protein extracts from hepatoma cell lines, HuH7 and FAO-1. However, only Cx32-B2 binding activity was detected in nuclear protein extract from normal rat liver tissue. This activity was significantly more abundant in rat liver tissue than in hepatoma cell lines and tissues from various other organs. By using methylation interference footprinting, the Cx32-B2 complex was localized to the region between mp -152 and -127 and a DNA probe containing this region bound to a 60-kDa protein in rat liver nuclear extracts. Mutation of two nucleotides in the Cx32-B2 binding site abrogated the formation of the Cx32-B2 protein-DNA complex and significantly reduced the transcriptional activity of the Cx32 promoter. These results indicate that the Cx32-B2 complex is an essential component of the rat Cx32 basal promoter and is likely a major factor in the preferential expression of this gene in the liver.

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