McStay, M.; Savage, K.; Stubbs, M.; Khan, K.; Dhillon, A.; Caplin, M.
April 2003
Gut;Apr2003 Supplement 1, Vol. 52, pA91
Academic Journal
Background: PTH-related protein (PTHrP) was identified in 1982 as the major factor responsible for humoral hypercalcaemia of malignancy HHM. Since then, it has been found to be expressed in malignancies such as breast and prostate cancer, which are not commonly associated with HHM, and has been found to regulate their growth. Preliminary studies have suggested a role for PTHrP in hepatocellular carcinoma. Aim: To assess the expression of PTHrP and its corresponding PTH/PTHrP type 1 receptor (PTH1R) in human hepatocellular carcinoma resection specimens and cell lines. Methods: Immunocytochemical Iocalisation of PTHrP and PTH1R was performed by the APAAP method on paraffin sections from 16 consecutive, pre-operatively eucalcaemic patients with well-defined hepatocellular carcinoma, and on the hepatocellular carcinoma cell lines, PLC/PRF/5, HepG2 and MCA RH 7777. Murine monoclonal antibodies to PTHrP(1-10) and PTH1R were used, and specificity was demonstrated by pre-absorbance of antibodies with epitope. Western immunoblotting, using the same anti-PTHrP antibody, was used to assess the expression of PTHrP by the cell lines. PTHrP(1-10) peptide was labelled with Alexa Fluor 488 and incubated with the cell lines for 1 hour at 37°C. Cells were counterstained with fluorescent nuclear stain and examined under fluorescence microscope with appropriate filters. Results: APl of the tumour specimens and cell lines showed positive cytoplasmic staining for PTHrP and PTH1R. Variable staining for both peptide and receptor was seen in the background liver of the tumour specimens, but in 11 cases the staining was stronger in the tumour. Cellular PTHrP was detected by Western blotting in all of the cell line extracts. Uptake of labelled PTHrP was seen in some cells in each of the cell lines examined. The pattern of uptake was predominantly cytoplasmic, but nuclear Iocalisation was also seen in some cells. Conclusion: PTHrP and PTH1R are expressed by hepatocellular carcinoma...


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