Analysis of alpha-fetoprotein gene expression in hepatocellular carcinoma and liver cirrhosis by in situ hybridization

We quantitated mRNA and protein for ornithine decarboxylase (ODC) and c-myc in formalin-fixed liver sections from 25 specimens of hepatocellular carcinoma (HCC) and seven normal livers by a non-radiolabeled in situ hybridization technique and immunohistochemistry. This non-radioactive in situ hybridization technique was highly specific, with virtually no background, and permitted quantitative analysis based on optical density. Reaction products were quantitated with computer-assisted microdensitometry. Samples were classified as normal, adjacent uninvolved, cirrhosis, well-differentiated HCC, and poorly-differentiated HCC. There was a progressive increase in all four parameters measured, ODC mRNA and protein, and c-myc mRNA and protein, from normal, to adjacent uninvolved liver, to cirrhosis, to well-differentiated HCC, to poorly-differentiated HCC. The sole exception was that ODC mRNA was lowest in cirrhosis. The patterns of ODC and c-myc gene expression are similar in HCC. The quantitative detection of ODC mRNA, c-myc mRNA, and their protein products in hepatocellular carcinoma and cirrhosis by in situ hybridization and immunohistochemical techniques may have a potential role in the study of hepatocarcinogenesis and in the diagnosis of hepatocellular carcinoma.

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In situ Hybridization: Application to the Study of Gene Expression during Experimental Hepatocarcinogenesis and Human Hepatocellular Carcinoma

Tumor Biology ◽

10.1159/000217652 ◽

1990 ◽

Vol 11 (4) ◽

pp. 173-180 ◽

Cited By ~ 4

Author(s):

D. Bernuau ◽

G. Feldmann

Keyword(s):

Gene Expression ◽

Hepatocellular Carcinoma ◽

In Situ Hybridization ◽

Human Hepatocellular Carcinoma

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Cellular analysis by in situ hybridization and immunoperoxidase of alpha-fetoprotein and albumin gene expression in rat liver during the perinatal period.

The Journal of Cell Biology ◽

10.1083/jcb.103.3.777 ◽

1986 ◽

Vol 103 (3) ◽

pp. 777-786 ◽

Cited By ~ 42

Author(s):

A M Poliard ◽

D Bernuau ◽

I Tournier ◽

L G Legrès ◽

D Schoevaert ◽

...

Keyword(s):

Gene Expression ◽

In Situ Hybridization ◽

Rat Liver ◽

Cdna Probe ◽

Relative Number ◽

Perinatal Period ◽

Alpha Fetoprotein ◽

Cellular Analysis ◽

Afp Mrna

To analyze at the cellular level the decrease in alpha-fetoprotein (AFP) gene expression during the early postnatal growth, we searched for AFP gene transcripts by in situ hybridization using a specific cDNA probe, and for the corresponding protein by immunocytochemistry, on rat liver sections at various times of the perinatal period. The relative number of mRNA sequences was evaluated by Northern blot analysis. Albumin (ALB) gene expression was studied simultaneously with the same techniques. In 17-19-d-old fetuses all hepatocytes express simultaneously, for both genes, the mRNAs and the corresponding proteins. During the first postnatal weeks, at a time when the global number of AFP mRNA molecules decreases, all hepatocytes still contain cytoplasmic transcripts and protein. A zonal heterogeneity in the level of AFP gene expression develops around the first week, a higher number of gene products being detected in perivenous than in periportal hepatocytes. This heterogeneity persists until the fourth week when AFP mRNA sequences and protein are barely detectable. All hepatocytes express the ALB gene after birth, but at around the second week, a periportal intensification of the in situ hybridization signal and immunostaining becomes apparent. Our data indicate that co-expression of the AFP and ALB genes by all hepatocytes is a normal step in liver ontogeny; the diminution of AFP gene expression after birth is not the result of the disappearance of specialized cell clones; and zonal quantitative differences in the level of AFP and ALB gene expression are observed within the maturing liver lobule.

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Comparative Genomic Hybridization of Hepatocellular Carcinoma: Correlation With Fluorescence In Situ Hybridization in Paraffin-embedded Tissue

Molecular Diagnosis ◽

10.2165/00066982-200106010-00004 ◽

2001 ◽

Vol 6 (1) ◽

pp. 27-37 ◽

Cited By ~ 17

Author(s):

UMA N. M. RAO ◽

SUSANNE M. GOLLIN ◽

STACIE BEAVES ◽

KATHLEEN CIEPLY ◽

MICHAEL NALESNIK ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

Comparative Genomic Hybridization ◽

Comparative Genomic ◽

Genomic Hybridization

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RNA In Situ Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of Drosophila Species

Methods and Protocols ◽

10.3390/mps4010020 ◽

2021 ◽

Vol 4 (1) ◽

pp. 20

Author(s):

Mujeeb Shittu ◽

Tessa Steenwinkel ◽

William Dion ◽

Nathan Ostlund ◽

Komal Raja ◽

...

Keyword(s):

Gene Expression ◽

In Situ Hybridization ◽

Expression Patterns ◽

Drosophila Species ◽

Gene Expression Patterns ◽

Probe Design ◽

Tissue Preparation ◽

Design And Synthesis ◽

Rna In Situ Hybridization

RNA in situ hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model Drosophila species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model Drosophila species.

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