Skip to main content
NCBI home page
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1992 Jan;45(1):21–24. doi: 10.1136/jcp.45.1.21

In situ hybridisation of albumin mRNA in normal liver and hepatocellular carcinoma with a digoxigenin labelled oligonucleotide probe.

G I Murray 1, P J Paterson 1, S W Ewen 1, W T Melvin 1
PMCID: PMC495803  PMID: 1311001

Abstract

AIMS: To study the localisation and distribution of albumin mRNA in normal liver and hepatocellular carcinoma by in situ hybridisation with an oligonucleotide probe. METHODS: A 51 base oligonucleotide was synthesised from a sequence at the 5' end of the human albumin gene and the probe was labelled at its 3' end with digoxigenin 11-dUTP. Formalin fixed, wax embedded sections of liver biopsy specimens were used to study the localisation and distribution of albumin mRNA. After in situ hybridisation the bound probe was visualised using a digoxigenin antibody conjugated with alkaline phosphatase. RESULTS: In normal liver albumin mRNA was detected in hepatocytes and no positive signal was observed in biliary epithelium, vascular endothelium, or Kupffer cells. In 75% (9/12) of the hepatocellular carcinomas studied a positive hybridisation signal was observed in tumour cells. CONCLUSIONS: Albumin mRNA can be detected in sections of formalin fixed, wax embedded liver, a digoxigenin labelled probe is ideally suited for in situ hybridisation of liver because there is no background from the detection system. The identification of albumin mRNA may be a useful marker of hepatocellular carcinoma, and the demonstration of albumin mRNA by in situ hybridisation overcomes the potential background problem associated with albumin immunohistochemistry.

Full text

PDF
21

Images in this article

22Image
on p.22
23Image
on p.23
23Image
on p.23

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akhtar N., Ruprai A., Pringle J. H., Lauder I., Durrant S. T. In situ hybridization detection of light chain mRNA in routine bone marrow trephines from patients with suspected myeloma. Br J Haematol. 1989 Nov;73(3):296–301. doi: 10.1111/j.1365-2141.1989.tb07743.x. [DOI] [PubMed] [Google Scholar]
  2. Beaune P. H., Umbenhauer D. R., Bork R. W., Lloyd R. S., Guengerich F. P. Isolation and sequence determination of a cDNA clone related to human cytochrome P-450 nifedipine oxidase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8064–8068. doi: 10.1073/pnas.83.21.8064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dirks R. W., van Gijlswijk R. P., Tullis R. H., Smit A. B., van Minnen J., van der Ploeg M., Raap A. K. Simultaneous detection of different mRNA sequences coding for neuropeptide hormones by double in situ hybridization using FITC- and biotin-labeled oligonucleotides. J Histochem Cytochem. 1990 Apr;38(4):467–473. doi: 10.1177/38.4.2108203. [DOI] [PubMed] [Google Scholar]
  4. Dugaiczyk A., Law S. W., Dennison O. E. Nucleotide sequence and the encoded amino acids of human serum albumin mRNA. Proc Natl Acad Sci U S A. 1982 Jan;79(1):71–75. doi: 10.1073/pnas.79.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Farquharson M., Harvie R., McNicol A. M. Detection of messenger RNA using a digoxigenin end labelled oligodeoxynucleotide probe. J Clin Pathol. 1990 May;43(5):424–428. doi: 10.1136/jcp.43.5.424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Feldmann G., Penaud-Laurencin J., Crassous J., Benhamou J. P. Albumin synthesis by human liver cells: its morphological demonstration. Gastroenterology. 1972 Dec;63(6):1036–1048. [PubMed] [Google Scholar]
  7. Grody W. W., Cheng L., Lewin K. J. In situ viral DNA hybridization in diagnostic surgical pathology. Hum Pathol. 1987 Jun;18(6):535–543. doi: 10.1016/s0046-8177(87)80353-2. [DOI] [PubMed] [Google Scholar]
  8. Herrington C. S., Burns J., Graham A. K., Evans M., McGee J. O. Interphase cytogenetics using biotin and digoxigenin labelled probes I: relative sensitivity of both reporter molecules for detection of HPV16 in CaSki cells. J Clin Pathol. 1989 Jun;42(6):592–600. doi: 10.1136/jcp.42.6.592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jirikowski G. F., Ramalho-Ortigao J. F., Lindl T., Seliger H. Immunocytochemistry of 5-bromo-2'-deoxyuridine labelled oligonucleotide probes. A novel technique for in situ hybridization. Histochemistry. 1989;91(1):51–53. doi: 10.1007/BF00501911. [DOI] [PubMed] [Google Scholar]
  10. Kojiro M., Kawano Y., Isomura T., Nakashima T. Distribution of albumin- and/or alpha-fetoprotein-positive cells in hepatocellular carcinoma. Lab Invest. 1981 Mar;44(3):221–226. [PubMed] [Google Scholar]
  11. Larsson L. I., Hougaard D. M. Optimization of non-radioactive in situ hybridization: image analysis of varying pretreatment, hybridization and probe labelling conditions. Histochemistry. 1990;93(4):347–354. doi: 10.1007/BF00315849. [DOI] [PubMed] [Google Scholar]
  12. Larsson L. I. In situ hybridization of messenger RNA sequences. Histochem J. 1989 Aug;21(8):435–440. doi: 10.1007/BF01845792. [DOI] [PubMed] [Google Scholar]
  13. Lewis M. E., Krause R. G., 2nd, Roberts-Lewis J. M. Recent developments in the use of synthetic oligonucleotides for in situ hybridization histochemistry. Synapse. 1988;2(3):308–316. doi: 10.1002/syn.890020321. [DOI] [PubMed] [Google Scholar]
  14. Lloyd R. V., Jin L., Fields K. Detection of chromogranins A and B in endocrine tissues with radioactive and biotinylated oligonucleotide probes. Am J Surg Pathol. 1990 Jan;14(1):35–43. doi: 10.1097/00000478-199001000-00004. [DOI] [PubMed] [Google Scholar]
  15. Morimoto H., Monden T., Shimano T., Higashiyama M., Tomita N., Murotani M., Matsuura N., Okuda H., Mori T. Use of sulfonated probes for in situ detection of amylase mRNA in formalin-fixed paraffin sections of human pancreas and submaxillary gland. Lab Invest. 1987 Dec;57(6):737–741. [PubMed] [Google Scholar]
  16. Samoszuk M., Nansen L. Detection of interleukin-5 messenger RNA in Reed-Sternberg cells of Hodgkin's disease with eosinophilia. Blood. 1990 Jan 1;75(1):13–16. [PubMed] [Google Scholar]

Articles from Journal of Clinical Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES