Insulin like growth factors and p62 c-myc in solid tumors
Main researcher
: ČABRIJAN, TOMISLAV (74820) Assistants
MISJAK, MIRA (68654)
ALAČ, MIRJANA (74796)
ŠUŠKOVIĆ, TOMISLAV (38976)
KRPAN, RUŽICA (145492)
KARAPANDŽA, NIKOLA (127095)
SILOBRČIĆ, IVO (94234)
FRKOVIĆ-GRAZIO, SNJEŽANA (175394)
PETEK, MARIJAN (67993)
ALTABAS, VELIMIR (900005)
LIPOVAC, MIRNA (900719)
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
Communication
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.
COOPERATION - PROJECTS
Name of project
: Ekspresija i uloga onkogena i faktora rasta u
malignim tumorima Name of institution: Institut "Ruđer Bošković" City: 10000 - Zagreb, Croatia
COOPERATION - INSTITUTIONS
Name of institution
: Unniversitaetskrankenhaus "Eppendorf" Type of institution: Economical/Production City: D-2000-52 - Hamburg, Njemačka