SVIBOR - Project code: 3-01-209


Strossmayerov trg 4, HR - 10000 ZAGREB
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Project code: 3-01-209

Insulin like growth factors and p62 c-myc in solid tumors

Main researcher: ČABRIJAN, TOMISLAV (74820)

Type of research: applied
Duration from: 01/01/91. to 01/01/96.

Papers on project (total): 48
Papers on project quoted in Current Contents: 4
Institution name: Bolnica "Sestre milosrdnice", Zagreb (134)
Department/Institute: Institut for Endocrinology, diabetes and diseases of metabolism Department of Internal medicine University Hospital "Sestre milosrdnice", Zagreb
Address: Vinogradska 29
City: 10000 - Zagreb, Croatia
Phone: 385 (0)17-46-66
Fax: 385 (0)17-24-53

Summary: The expression of insulin-related growth factors (insulinoids), detectable by specific radioimmunoassay or immunohistochemical staining, was observed in the tumor tissue removed surgically from patients suffering from solid tumors. Among different insulinoids, insulin-like growth factor (IGF-I), nerve growth factor (NGF), insulin like substances, and insulin were observed in several urinary bladder, rectal, anal, and breast carcinomas, as well as in melanomas and myxoma. Both, high preoperative insulinoid and low blood glucose concentrations returned to normal level after surgery. According to molecular weight (SDS-PAGE, chromatography) and cross-reactivity with specific antibodies, these substances may be classified in three groups:120-150 kDa, 40 kDa, 6-10 kDa. Normally, oncoprotein p62 c-myc is localized in the cell nucleus. In freshly frozen sections, normal immunohistochemical localization of this antigen is preserved but some morphological details have been hampered by destruction of cell and tissue architecture during cryosectioning.Previously we have shown that in paraffin embedded sections the commonly observed cytoplasmic staining for p62 c-myc resulted from inadequate fixation. We found that a more adequate fixation procedure included treatment with cold acetone (-20 oC) followed by methyl benzoate and xylene (AMeX). We wished to eliminate artifacts of cryosectioning and fixation in formalin. In this study we evaluated the AMeX procedure in immunohistochemical localization of p62 c-myc in infiltrating ductal breast carcinoma tissue freshly frozen, AMeX treated and then embedded in paraffin (FF-AMeX). To delineate the distribution and intensity of staining for p62 c-myc in tumor cells, we used the monoclonal antibody 155-11C7 raised against a synthetic myc peptide. We assessed the intensity of immunostaining and percentage of positive tumor cells and observed more intense staining of p62 c-myc in a freshly frozen section than in FF-AMeX sections, but tissue and cell architecture were much better preserved i FF-AMeX. Our results show that the AMeX procedure in previously frozen tissues resulted in reproducible nuclear immunostaining for p62 c-myc. This finding is important because tissues often kept frozen for steroid receptor analysis can be used for immunohistochemical localization of oncoproteins.

Keywords: insulin like growth factors, oncogens, p62 c-myc, immunohistochemical localization of oncoproteins, solid tumors, insulinoma,oncogenes,point mutation,

Research goals: The goal of proposed examinations is the detection of insulinlike substances (insulinoids), proteins, and their carriers inthe serum of patients with solid breast, thyreoidea, pancreas andgastrointestinal tract tumors.We also want to follow expressionof p62 c-myc oncoprotein by immunohistochemical methods in frozensolid tumor tissues embedded in parafin. That is very simple andusefull detecting method for this oncoprotein, and provides theusing of frozen tumor tissue samples (for example breast tumortissue), which were alredy stored for steroid receptor analysis.Insulin and insulin-like substances, such as p62 c-mycexpression will be compared with histological type, size, thelevel of malignacy, presence of local metastases, clinical andlaboratory findings. That would be very useful for prognosis andtheraphy of those tumors.Special attention will be given to therelation of insulinoids and c-myc oncoproteins.


  1. Name of project: Ekspresija i uloga onkogena i faktora rasta u malignim tumorima
    Name of institution: Institut "Ruđer Bošković"
    City: 10000 - Zagreb, Croatia


  1. Name of institution: Unniversitaetskrankenhaus "Eppendorf"
    Type of institution: Economical/Production
    City: D-2000-52 - Hamburg, Njemačka

  2. Name of institution: Institut "Ruđer Bošković"
    Type of institution: University/Faculty
    City: 10000 - Zagreb, Croatia

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Last update: 10/02/95