Immunohistochemical markers of cancerogenesis in the lung
Walentyn Pankiewicz1,4, Łukasz Minarowski1, Wiesława Niklińska2,
Wojciech Naumnik3,
Jacek Nikliński4 and Lech Chyczewski1
Departments of: 1Clinical Molecular Biology, 2Histology and Embryology,
3Lung Diseases and Tuberculosis and 4Thoracic Surgery, Medical University of Białystok, Poland
Abstract: Lung cancer is the leading cause of cancer deaths for people of both sexes worldwide. Early diagnosis of precancer
lesions may be of crucial significance to lowering lung cancer mortality. The World Health Organization has defined
three preneoplastic lesions of the bronchial epithelium: squamous dysplasia and carcinoma in situ, atypical adenomatous
hyperplasia and diffuse idiopathic pulmonary neuroendocrine cell hyperplasia. These lesions are believed to progress to
squamous cell carcinoma, adenocarcinoma and carcinoid tumors, respectively. Apart from WHO classification, two other
lesions such as bronchiolization and bronchiolar columnar cell dysplasia (BCCD) can be observed and thought to be preneoplastic
lesions leading to adenocarcinoma. In this review we summarize the data of morphological and cell cycle related
proteins changes in both central and peripheral compartments of lung. Many molecular changes, which accompany the
multistep process of the development of invasive types of cancer, may be observed thanks to the application of immunohistochemical
markers. A deeper knowledge of molecular and genetic changes accompanying pre-cancer states may show
new directions of early diagnostics of cancer development.
Author’s e-mail: wal@amb.edu.pl
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Basal cell subpopulation as putative human prostate carcinoma stem cells
Aleksandar Kuzmanov1, Soren Hayrabedyan1, Milen Karaivanov2, Krassimira Todorova1
1Institute of Biology and Immunology of Reproduction, Bulgarian Academy of Sciences, Sofia, and
2Department of Pathology, University Hospital, Pleven, Bulgaria
Abstract: The present study examines the expression of p63, glutathione S-transferase-? (GSTP1) and ?-methylacyl-CoAracemase
(AMACR) in serial slices in proliferative inflammatory atrophy (PIA) in order to implicate that some of the basal
cells are probably the putative human prostate carcinoma stem cells (PHPCSC). Archived tissue sections obtained after radical
prostatectomy from cases (n=30) comprising of PIA were tested using immunohistochemistry with antibodies against
AMACR (Dako), p63 and GSTP1 (Labvision) and visualized by biotin-streptavidin-peroxidase kit (DAKO LSAB 2 kit).
Quantitative immunohistochemistry analysis (QIHC) of the studied antigen expression levels revealed that there are two
populations of p63 basal cells. Type I basal cells had high AMACR, low GSTP1 and p63 expression. Type II basal cells had
low AMACR, high GSTP1 and p63 expression. Therefore, we propose that the putative human prostate carcinoma stem cells
probably reside within the population of type I basal cells.
Author’s e-mail: krasiot@abv.bg
STAT3, HIF-1α, EPO and EPOR - signaling proteins in human primary ductal breast cancers
Andrzej Wincewicz, Mariola Sulkowska, Mariusz Koda, Tomasz Leśniewicz, Luiza Kanczuga-Koda and Stanisław Sulkowski
Departments of Clinical and General Pathomorphology, Medical University of Białystok, Poland
Abstract: STAT3 upregulates expression of HIF-1 induced EPO. Receptor EPOR was reported to activate STAT3. Our
study was aimed at demonstration of tissue immunoreactivities of those proteins and determination of their relationships in
reference to clinicopathological variables of breast cancers. We detected STAT3, HIF-1α, EPO and EPOR in specimens of
76 human, female, ductal breast cancers by immunohistochemistry. STAT3 was detected in 38 of 76 cancers (50%). HIF-1?
was found in 55 cases (72%). EPO positive tumors comprised 89% of all the cancers (68 cases). EPOR was also visualized
in 55 cases (72%). Anti-HIF-1α and anti-STAT3 stained nuclei and cytoplasm of breast cancer cells in diffuse and finely
granular fashion. Strong membranous expressions of EPO and EPOR were distributed in cytoplasmic and membranous
granularity or diffuse staining. STAT3 correlated with HIF-1 in general (r=0.4012, p<0.0001) and in different patients' subgroups.
STAT3 was significantly associated with EPO and EPOR in all the cancers (r=0.2370, p=0.039 and r=0.3336,
p=0.003, respectively). Besides a correlation between STAT3 and EPOR in node negative ones, STAT3 wasn't related to
EPO and EPOR in remaining subgroups. HIF-1α correlated with EPO and EPOR in most of analyzed groups. Immunoreactivity
to EPO generally was associated with EPOR (r=0.3520, p=0.002). Statistically analyzed distributions of the proteins
reflected functional dependences among STAT3, HIF-1α, EPO and EPOR in cellular signal conduction.
Author’s e-mail: sulek@zeus.amb.edu.pl
Expression of FAS/APO 1/CD 95 in thyroid tumors
Marta Rzeszutko1, Wojciech Rzeszutko1, Piotr Dzięgiel2, Waldemar Balcerzak3,
Krzysztof Kaliszewski3, Marek Bolanowski4
Departments of: 1Pathological Anatomy, 2Histology and Embryology, 3General and Endocrinological
Surgery and 4Endocrinology and Diabetology, University Medical School, Wrocław, Poland
Abstract: Using immunohistochemistry, Fas/Apo-1 protein expression was investigated in thyroid cancers of 67 patients.
Thyroid biopsies from twenty eight patients with benign thyroid diseases were also examined. The patients with thyroid cancer
manifested a variable histology of the cancer, including 14 patients with follicular carcinoma, 48 with papillary carcinoma,
5 patients with medullary carcinoma. The benign thyroid disease involved nodular goitre in 11 patients and follicular
adenoma in other 17 patients. The study aimed at examining immunohistochemical expression of Fas protein in order to
determine whether the level of its expression correlated with histological diagnosis. In individual patients Fas expression
was more prevalent in thyroid carcinomas as compared to benign tumors (p=0.001). A marked increase in Fas expression
was found in papillary carcinoma, as compared to follicular and medullary carcinomas (p=0.02). In conclusion, Fas was significantly
more frequently overexpressed in thyroid cancer, indicating its role in thyroid tumorigenesis.
Author’s e-mail: piotr@hist.am.wroc.pl
Cytosolic superoxide dismutase activity after photodynamic therapy, intracellular distribution
of Photofrin II and hypericin, and P-glycoprotein localization in human colon adenocarcinoma
Jolanta Saczko1, Julita Kulbacka1, Agnieszka Chwilkowska1, Andrzej Pola2, Mateusz Lugowski1,
Anna Marcinkowska1, Anna Malarska1, Teresa Banas1
1Department of Medical Biochemistry, 2Department of Biophysics, Wrocław Medical University, Wrocław, Poland
Abstract: In photodynamic therapy (PDT), a tumor-selective photosensitizer is administered and then activated by exposure
to a light source of applicable wavelength. Multidrug resistance (MDR) is largely caused by the efflux of therapeutics
from the tumor cell by means of P-glycoprotein (P-gp), resulting in reduced efficacy of the anticancer therapy. This study
deals with photodynamic therapy with Photofrin II (Ph II) and hypericin (Hyp) on sensitive and doxorubicin-resistant colon
cancer cell lines. Changes in cytosolic superoxide dismutase (SOD1) activity after PDT and the intracellular accumulation
of photosensitizers in sensitive and resistant colon cancer cell lines were examined. The photosensitizers' distributions indicate
that Ph II could be a potential substrate for P-gp, in contrast to Hyp. We observed an increase in SOD1 activity after
PDT for both photosensitizing agents. The changes in SOD1 activity show that photodynamic action generates oxidative
stress in the treated cells. P-gp appears to play a role in the intracellular accumulation of Ph II. Therefore the efficacy of
PDT on multidrug-resistant cells depends on the affinity of P-gp to the photosensitizer used. The weaker accumulation of
photosensitizing agents enhances the antioxidant response, and this could influence the efficacy of PDT.
Author’s e-mail: michal@bioch.am.wroc.pl
Apoptosis in ovarian cells in postmenopausal women
Agnieszka Brodowska1, Maria Laszczyńska2, Andrzej Starczewski1
1Department of Reproduction and Gynecology, 2Laboratory of Embryology, Pomeranian Medical University, Szczecin, Poland
Abstract: Apoptosis is a natural process which accompanies human ovary from the moment of birth until old age. While it
is a well-known process at the reproductive age, it still needs to be thoroughly examined when referring to the postmenopausal
age. The study involved 30 postmenopausal women who had their ovaries removed by laparotomy due to nonneoplastic
diseases of the uterus. The women were divided into 3 groups depending on the time that had passed since the
last menstruation. Group A consisted of women who had their last menstruation no more than 5 years earlier. In group B
menopause occurred 5 to 10 years earlier. Group C was composed of patients who had the last menstruation over 10 years
earlier. In all the patients concentrations of follitropin (FSH) and estradiol (E2) in blood plasma were measured. Ovarian tissue
was obtained during surgery. For morphological studies, ovaries were fixed in Bouin`s solution and 4% formalin and
embedded in paraffin. Morphological analysis was carried out after hematoxylin-eosin (H-E) staining. For histochemical
detection of apoptotic cells (in situ localization of fragment DNA), the TUNEL method was used. The expression of caspase-3
positive cells was determined immunohistochemically in paraffin-embedded specimens. Comparing to groups A and
B, the ovaries in group C contained small number of corpora albicantia located in the medullary part as well as thinned blood
vessels and few lymphatic vessels and nerves. In contrast to group A where the number of TUNEL-positive cells was high
and caspase-3 expression was observed, no TUNEL-positive nuclei and caspase-3 expression were found in the examined
ovaries of group C women.
Author’s e-mail: laszcz@sci.pam.szczecin.pl
Oral β-glucan adjuvant therapy converts nonprotective Th2 response to protective Th1 cell-mediated immune
Author’s e-mail: mibaran@cyf-kr.edu.pl
Alteration of parafollicular (C) cells activity in the experimental model of hypothyroidism in rats
Neuroendocrine cells in the nasal mucosa - preliminary report
Localization of RNA transcription sites in insect oocytes using microinjections of 5-bromouridine 5’-triphosphate
Dmitry Bogolyubov
Institute of Cytology, Russian Academy of Sciences, St. Petersburg, Russia
Abstract: In the present study we used 5-bromouridine 5'-triphosphate (BrUTP) microinjections to localize the transcription
Vitreous humour as a potential DNA source for postmortem human identification
Ireneusz Sołtyszewski1, Anna Niemcunowicz-Janica2, Witold Pepiński2, Magdalena Spólnicka2, Renata Zbiec3, Jerzy Janica2
1Department of Criminalistics and Forensic Medicine, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland
Abstract: Purpose: The aim of this study was the assessment of vitreous humor as a potential DNA for forensic human postmortem
response in mammary tumor-bearing mice
Jarek Baran1,2, Daniel J. Allendorf1, Feng Hong1, Gordon D. Ross1
1Tumor Immunobiology Program, James Graham Brown Cancer Center, University of Louisville, Lousville, KY, USA
2Department of Clinical Immunology, Polish-American Institute of Pediatrics, Jagiellonian University Medical College, Cracow, Poland
Abstract: Beta (1-3)-D-glucans were identified almost 40 years ago as biological response modifiers that stimulated tumor
rejection. In vitro studies have shown that β-glucans bind to a lectin domain within complement receptor type 3 (CR3), or
to, more recently described dectin-1 a β-glucan specific receptor, acting mainly on phagocytic cells. In this study, we
assessed the intracellular cytokine profiles of peripheral blood lymphocytes from mice bearing mammary tumors receiving
i.v. anti-tumor mAbs combined or not with whole glucan particle suspension given orally (WGP, 400 μg every 24 hours).
The proportions of T cells producing IL-4 and IFNγ were determined by flow cytometry. The proportion of T cells producing
IL-4 was significantly higher in tumor-bearing mice not receiving β-glucan-enhanced therapy. Conversely, T cells from
mice undergoing β-glucan-enhanced therapy showed increased production of the Th1 cytokine IFNγ. The switch from a Th2
to a Th1 response after WGP therapy was possibly mediated by intestinal mucosal macrophages releasing IL-12.
Robert Łukasz Zbucki1, Maria Malgorzata Winnicka1, Boguslaw Sawicki2, Beata Szynaka3,
Anna Andrzejewska3, Zbigniew Puchalski4
1Department of General and Experimental Pathology, 2Department of Histology and Embryology,
3Department of Medical Pathomorphology, 41st Department of General and Endocrinological Surgery,
Medical University of Białystok, Poland
Abstract: Our previous study has shown the alteration of C cells activity in rats with experimental model of hyperthyroidism.
The aim of the present study was the evaluation of parafollicular cells activity in rats with hypothyroidism evoked
by propylthiouracil (PTU) given in drinking water over 21 days. Histological, ultrastructural and immunocytochemical studies
using specific antibodies against calcitonin and CGRP were performed on thyroid glands taken from experimental and
control groups of rats. Moreover, in all animals the calcitonin plasma levels were evaluated by radioimmunoassay. After
chronic administration of PTU, thyroid image showed predominant microfollicular hyperplasia and attenuated density of
parafollicular cells. The intensity of immunocytochemical reactions for CT and CGRP were weaker in the majority of C
cells in comparison to the control rats, in which strong immunocytochemical reaction was observed. Examination in the
electron microscope reveals the features of hypoactivity both in follicular and parafollicular cells, in which the quantity and
electron density of secretory granules were smaller in comparison to the control group. These microscopic changes were
accompanied by a significant decrease of calcitonin plasma concentration. Alteration of C cells activity in the experimental
model of hypothyroidism, accompanied by microfollicular hypertrophy, may point to the mutual cooperation between
parafollicular and follicular cells.
Andrzej Sieśkiewicz1, Monika Olszewska2, Ewa Olszewska1, Magdalena Garbowicz3, Sylwia Trojan3, Lech Chyczewski3, Marek Rogowski1
Departments of: 1Otolaryngology Head and Neck Surgery, 2Emergency Medicine and 3Clinical Molecular
Biology, Medical University of Białystok, Białystok, Poland
Abstract: The role of neuroendocrine cells (NC) in physiology and pathology of the human's respiratory tract is not fully
understood. The aim of the study was the quantitative and morphometric assessment of NC in nasal mucosa in some pathological
states. Patients and method: 40 patients, aged 28-63 years, with clinical signs of chronic, hypertrophic rhinosinusitis
were qualified for the study. Rhinitis chronica hypertrophica coexisted with aspirin triad or asthma in 10 patients (group
I), with advanced obstructive sleep apnea syndrome (OSAS) in 10 patients (respiratory disturbance index, RDI>40, group
II). Group III consisted of 10 patients with simple rhinitis chronica hypertrophica who habitually smoked cigarettes (at least
20 cigarettes a day) while 10 non-smoking patients with simple rhinitis chronica hypertrophica were qualified to the control
group. Fragments of nasal mucosa of approximately 0.5 cm2 were collected from medial or inferior turbinate during
mucoplasty procedures. NC were detected immunohistochemically using antibodies against chromogranin A (DAKO). The
microscopic sections were evaluated in the light microscopy. Results: The study did not reveal the increased number of NC
in examined fragments of nasal mucosa. Scattered NC were detected in single preparations of nasal mucous membrane in
some patients in all groups. The number of detected neuroendocrine cells did not differ statistically between groups.
Author’s e-mail: sieska@interia.pl
sites in oocytes of insects with different types of the ovarium structure: panoistic, meroistic polytrophic, and meroistic
telotrophic. We found that in an insect with panoistic ovaries (Acheta domesticus), oocyte nuclei maintain their transcription
activity during the long period of oocyte growth. In insects with meroistic ovaries (Tenebrio molitor and Panorpa communis),
early oocyte chromosomes were found to be transcriptionally active, and some transcription activity still persist
while the karyosphere, a compact structure formed by all condensed oocyte chromosomes, begins to develop. At the latest
stages of karyosphere development, no anti-Br-RNA signal was registered in the karyosphere.
Author’s e-mail: dmitr@mail.cytspb.rssi.ru
2Department of Forensic Medicine, Medical University of Białystok, Białystok, Poland
3Department of Biology, Central Forensic Laboratory of Police, Warsaw, Poland
identification. Material and methods: Vitreous humor samples were collected using two alternative approaches from
25 corpses of either sex during autopsies. DNA was extracted by standard organic method. Recovered DNA was quantitiated
fluorometrically. AmpFlSTR SGM Plus kit and ABI 310 Genetic Analyzer (Applera) were used to obtain genetic profiles.
Results: Different DNA yields were quantitated in vitreous body depending on cause of death and sampling approach.
Conclusion: Vitreous humor is a potential DNA for forensic human postmortem identification depending on a sampling
method used.
Author’s e-mail: pepinski@amb.edu.pl