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1.
Pathol Oncol Res ; 21(4): 921-7, 2015 Sep.
Article in English | MEDLINE | ID: mdl-25743258

ABSTRACT

Prostate cancer is the second most common malignancy in men worldwide. Abnormal epigenetic alterations such as DNA methylation and histone modification play an important role in tumor initiation, progression and regulation of cancer-related genes such as integrin α4 and E-cadherin. Expression of these genes was determined by semi-quantitative reverse transcriptase-PCR in prostate cancer cell lines, DU145 and PC3, before and after treatment with 5-aza-2-deoxycytidine and trichostatin A. Laser capture microdissection microscopy was used to obtain exclusively affected epithelial cells from prostate gland biopsies of 30 patients with prostate cancer and 40 with benign prostate hyperplasia. DNA bisulfite modifications followed by methylation-specific PCR were used to evaluate the promoter methylation status of E-cadherin and α4 integrin genes in extracted DNA from patients and aforementioned cell lines. The integrin α4 promoter in DU145 was fully methylated, whereas in PC3 cells, partial methylation was detected. E-cadherin was expressed in both cell lines; trichostatin A and 5-aza-2-deoxycytidine treatment had no effect on E-cadherin expression, however the combined treatment of both drugs or 5-aza-2-deoxycytidine alone increased integrin α4 expression. Integrin α4 and E-cadherin were hypermethylated in 66.6 % and 6.6 % of prostate cancer cases, respectively; no hypermethylation was observed in patients with benign prostate hyperplasia. These results together suggest that aberrant DNA methylation is one of the mechanisms involved in integrin α4 expression and may play an important role in human prostate carcinogenesis. In addition, the higher rate of integrin α4 gene methylation in prostate cancer patients elects it as a potential molecular tumor marker.


Subject(s)
Cadherins/genetics , DNA Methylation , Gene Expression Regulation, Neoplastic/drug effects , Integrin alpha4/genetics , Prostatic Hyperplasia/genetics , Prostatic Neoplasms/genetics , Antimetabolites, Antineoplastic/pharmacology , Azacitidine/analogs & derivatives , Azacitidine/pharmacology , Biomarkers, Tumor/genetics , CpG Islands , Decitabine , Histone Deacetylase Inhibitors/pharmacology , Humans , Hydroxamic Acids/pharmacology , Male , Promoter Regions, Genetic/genetics , Prostatic Hyperplasia/drug therapy , Prostatic Hyperplasia/pathology , Prostatic Neoplasms/drug therapy , Prostatic Neoplasms/pathology , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Tumor Cells, Cultured
2.
Mol Biol Rep ; 41(5): 2835-44, 2014 May.
Article in English | MEDLINE | ID: mdl-24452720

ABSTRACT

Allelic variation of BAT-25 (a 25-repeat quasimonomorphic poly T) and BAT-26 (a 26-repeat quasimonomorphic polyA) loci as two mononucleotide microsatellite markers, were analyzed with high-performance liquid chromatography (HPLC) compared with Real-Time PCR using hybridization probes. BAT-26 and BAT-25 markers were used to determine an appropriate screening technique with high sensitivity and specificity to diagnose microsatellite instability (MSI) status in patients with colorectal cancer (CRC). One of the pathways in colorectal tumor genesis is microsatellite instability (MSI+). MSI is detected in about 15% of all CRCs; 3% are of these are associated with Lynch syndrome and the other 12% are caused by sporadic. Colorectal tumors with MSI have distinctive features compared with microsatellite stable tumors. Due to the high percentage of MSI+ CRC in Iran, screening of this type of CRC is imperative. Two markers were analyzed in tissues and sera of 44 normal volunteers and tumor and matched normal mucosal tissues as well as sera of 44 patients with sporadic CRC. The sensitivity and specificity of BAT-26 with real time PCR method (Hybridization probe) were 100% in comparison with sequencing method as the gold standard, while HPLC had a lower sensitivity and specificity. According to HPLC data, BAT-26 was more sensitive than BAT-25 in identifying MSI tumors. Therefore, MSI typing using the BAT-26 hybridization probe method compared to HPLC could be considered as an accurate method for diagnosing MSI in CRC tumors but not in serum circulating DNAs.


Subject(s)
Colorectal Neoplasms/genetics , Microsatellite Instability , Microsatellite Repeats , Adult , Aged , Biomarkers, Tumor , Case-Control Studies , Cell Line, Tumor , Chromatography, High Pressure Liquid , Colorectal Neoplasms/pathology , Female , Genetic Markers , Germ-Line Mutation , Humans , Male , Middle Aged , Neoplasm Grading , Neoplasm Staging , Nucleic Acid Hybridization , ROC Curve , Real-Time Polymerase Chain Reaction
3.
Mol Biol (Mosk) ; 47(5): 787-95, 2013.
Article in Russian | MEDLINE | ID: mdl-25509351

ABSTRACT

Reports on the isolation of mesencnymal stromal cells (MSCs) from granulocyte colony stimulating factor mobilized peripheral blood (G-CSF-mobilized PB) using regular culturing techniques are controversial. Enrichment techniques such as CD133 isolation have increased the success rates. CD271 is a well-known marker for enrichment of MSCs from bone marrow (BM). In the present study, we aimed to find out whether CD271 enrichment can help isolation of MSCs from G-CSF-mobilized PB. Five G-CSF-mobilized PB samples were collected from the remnant parts of the bags used for BM transplantation. Five BM samples were used as the control. Mononuclear cells (MNCs) from both resources were collected and underwent magnetic sorting for CD271-positive cells. The isolated cells were cultured, undergoing flowcytometry and differentiation assays to determine if they fulfill MSCs characteristics. CD271-positive portion of G-CSF-mobilized PB did not yield any cell outgrowth but the BM counterpart could successfully form MSC colonies. Although the percentage of CD271+ cells showed no difference between BM-MNCs and G-CSF-mobilized PB-MNCs, hematopoietic markers such as CD45, CD34 and CD133 composed a higher percentage of CD271-positive cells in the G-CSF-mobilized PB group. Results obtained indicated that CD271 enrichment does not help isolation of MSCs from G-CSF-mobilized PB. In this source, almost all of the CD271+ cells are from hematopoietic origin and the frequency of MSCs is so low that possibly during the process of cell isolation most of them are lost and the isolation fails.


Subject(s)
Bone Marrow Cells/cytology , Cell Culture Techniques , Mesenchymal Stem Cells/cytology , Nerve Tissue Proteins/genetics , Receptors, Nerve Growth Factor/genetics , Bone Marrow Cells/metabolism , Cell Differentiation/genetics , Flow Cytometry , Granulocyte Colony-Stimulating Factor/genetics , Granulocyte Colony-Stimulating Factor/metabolism , Humans , Leukocytes, Mononuclear , Mesenchymal Stem Cells/metabolism , Nerve Tissue Proteins/metabolism , Receptors, Nerve Growth Factor/metabolism
5.
Cytopathology ; 18(3): 151-6, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17573764

ABSTRACT

OBJECTIVE: To study fine needle aspiration (FNA) cytological findings of splenic lesions and assess the role of FNA in the diagnosis of splenomegaly or splenic tumours. METHODS: This study consisted of 48 cases, 25 males and 23 females. The ages ranged between 3 and 71 years. Most of these cases were aspirated under ultrasonographic guidance and a small number were also aspirated directly by using a 22- to 23-gauge needles. The smears were stained with Wright-Giemsa and Papanicolaou methods. Special stains were used whenever necessary. RESULTS: In this study 14 cases were diagnosed as lymphoma-leukaemia, 7 cases as tuberculosis, 12 cases as kala-azar, 2 cases as hydatid cyst, 3 cases as storage diseases, 3 cases as simple cyst, 2 cases as myeloproliferative disorders, 2 cases as malignant tumours and 3 cases as hamartomas (these were wrongly diagnosed as malignant tumours). CONCLUSION: Splenic aspiration is a safe procedure and is very useful in the diagnosis of parasitic and infectious diseases, especially in endemic countries like Iran.


Subject(s)
Biopsy, Fine-Needle/methods , Spleen/pathology , Splenic Diseases/pathology , Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Reproducibility of Results
6.
Cancer Genet Cytogenet ; 165(2): 114-20, 2006 Mar.
Article in English | MEDLINE | ID: mdl-16527605

ABSTRACT

CTLA4 is a coinhibitory molecule expressed mainly on activated T lymphocytes. To test the putative involvement of CTLA-4 in inhibitory state of immunity to breast cancer, we genotyped 283 patients and 245 healthy control subjects for -1722 T/C, -1661 A/G, and -318 C/T single nucleotide polymorphisms in the promoter region of the CTLA4 gene. There were no significant differences in genotype, allele, or haplotype frequencies in all three loci between patients and healthy controls. Moreover, the incidence of the most frequent haplotype combination (TAC/TAC, T -1722, A -1661, C -318) was only slightly higher among healthy controls than patients (68.4 vs. 64.8%, P = 0.2). This haplotype combination was associated with lower stages of the disease (P = 0.0007), however, and higher estrogen receptor (ER) expression in patients (P = 0.006). Association with tumor prognostic or predictive factors was also observed with certain genotypes: the -1661 AA genotype was associated with lesser lymph node (LN) involvement (P = 0.017) and higher ER expression (P = 0.004), and the -318 CC genotype with lesser LN involvement (P = 0.007). These results suggest that CTLA4 promoter variants participate in the progression of breast cancer rather than in its initial development.


Subject(s)
Antigens, Differentiation/genetics , Breast Neoplasms/genetics , Promoter Regions, Genetic , Adult , Aged , Aged, 80 and over , Alleles , Antigens, CD , Breast Neoplasms/immunology , CTLA-4 Antigen , Female , Gene Frequency , Genotype , Humans , Middle Aged , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length
7.
Acta Cytol ; 44(4): 543-6, 2000.
Article in English | MEDLINE | ID: mdl-10934946

ABSTRACT

OBJECTIVE: To study the prevalence and significance of bull's eye (target) inclusions in neoplastic cells in malignant serous effusions. STUDY DESIGN: We reviewed malignant pleural, peritoneal and pericardial effusions from 289 patients who had proven cancer at known primary sites. The ages of the patients ranged from 5 to 72 years; 166 were male and 123 female. RESULTS: Bull's eye inclusions are an uncommon finding and appeared in only 13 cases of metastatic adenocarcinoma of the breast, stomach, colon, lung, ovary, pancreas and urinary bladder. They were positively stained with periodic acid-Schiff stain with diastase. The inclusions were not seen in cells of nonadenocarcinomatous neoplasms, such as squamous cell carcinoma, oat cell (small cell) carcinoma, neuroblastoma, lymphoma and germ cell tumors. CONCLUSION: Bull's eye inclusions are found in about 5% of malignant serous effusions containing cells of metastatic adenocarcinoma. The primary site of an adenocarcinoma cannot be deduced on the basis of the presence of inclusions.


Subject(s)
Adenocarcinoma/pathology , Ascitic Fluid/pathology , Inclusion Bodies , Pericardial Effusion/pathology , Pleural Effusion, Malignant/pathology , Adolescent , Adult , Aged , Biopsy , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Predictive Value of Tests
8.
Acta Cytol ; 44(3): 437-41, 2000.
Article in English | MEDLINE | ID: mdl-10834007

ABSTRACT

BACKGROUND: Cystic lesions of the brain may have diverse etiologies, ranging from true cysts to malignant tumors with cystic degeneration. Preoperative determination of the exact nature of them as well as intraoperative diagnosis may be sometimes difficult or even impossible. Sensitivity and specificity of diagnosis will be improved by introducing new methods or combining traditional procedures. CASES: Three metastatic brain carcinomas with primary sites of breast, pancreas and prostate presented as cystic lesions and were confused clinically with abscess. Intraoperative frozen section was not revealing. Cytologic study of sediments of aspirated fluid uncovered malignant cells. CONCLUSION: When combined with frozen section, intraoperative cytologic studies in the form of crush preparation, fine needle aspiration or evaluation of aspirated fluid in cystic lesions (as in our cases) can improve diagnostic accuracy by detecting important diagnostic features that otherwise may be missed.


Subject(s)
Brain Abscess/diagnosis , Brain Neoplasms/diagnosis , Brain Neoplasms/secondary , Breast Neoplasms/pathology , Diagnostic Errors , Monitoring, Intraoperative , Pancreatic Neoplasms/pathology , Prostatic Neoplasms/pathology , Adult , Breast Neoplasms/diagnostic imaging , Cytodiagnosis , Female , Humans , Male , Middle Aged , Neoplasm Metastasis , Tomography, X-Ray Computed
9.
Acta Cytol ; 44(2): 181-4, 2000.
Article in English | MEDLINE | ID: mdl-10740604

ABSTRACT

OBJECTIVE: To describe some rare cytologic findings in medullary thyroid carcinoma. STUDY DESIGN: Review of the fine needle aspiration smears from 15 cases of medullary thyroid carcinoma that were confirmed on histologic sections. The ages ranged between 31 and 67 years; 10 were female and 5 male. Thirteen were sporadic forms, and two were familial forms. RESULTS: Eight cases were classified as pleomorphic cell type and seven as monomorphic cell type. The smears revealed round, oval, triangular, polygonal and spindle-shaped cells, intracytoplasmic red granules, occasional intranuclear inclusions, amyloid, binucleated and multinucleated cells, and thick, granular chromatin. CONCLUSION: The rare cytologic findings in this study were grape cells, cytoplasmic nippling, elongated cytoplasmic processes, carrot-shaped nuclei, nuclear buddings, mast cell-like cells and a Burkitt's lymphoma-like appearance. These findings were rarely reported before.


Subject(s)
Carcinoma, Medullary/pathology , Thyroid Neoplasms/pathology , Adult , Aged , Biopsy, Needle , Cell Nucleus/pathology , Cytoplasm/pathology , Female , Humans , Male , Middle Aged
11.
Acta Cytol ; 43(5): 767-70, 1999.
Article in English | MEDLINE | ID: mdl-10518128

ABSTRACT

OBJECTIVE: To study the cytologic findings of papillary breast carcinoma by fine needle aspiration. STUDY DESIGN: The study group consisted of fine needle aspiration (FNA) specimens of breast tumors from nine patients performed during the period 1988-1997. Eight were female, and one was male. The FNA results were compared with the final histologic diagnosis. RESULTS: The tumor sizes were 4-6.5 cm. The aspirations yielded a good amount of bloody material. The smears revealed high cellularity, papillary clusters, isolated low-to-tall columnar cells, mild to moderate atypia, hemorrhagic background, foam and hemosiderin-laden macrophages, calcification, rare mitoses, palisading row of cells and bipolar cytoplasmic eosinophilic granules. The smears were diagnosed as either suspicious or suggestive of papillary carcinoma. The histologic examination revealed invasive papillary carcinoma. CONCLUSION: Papillary carcinoma of the breast can be diagnosed by using a panel of cytologic findings that includes hypercellularity, papillary clusters, hemorrhagic background, palisading rows of tall columnar cells, cellular atypia and calcification. The interesting finding in this study was the presence of eosinophilic bipolar cytoplasmic granules, which has not been reported before.


Subject(s)
Breast Neoplasms, Male/pathology , Breast Neoplasms/pathology , Carcinoma, Papillary/pathology , Aged , Biopsy, Needle/methods , Breast Neoplasms/surgery , Breast Neoplasms, Male/surgery , Carcinoma, Papillary/surgery , Female , Humans , Male , Mastectomy, Segmental , Middle Aged
12.
Acta Cytol ; 43(2): 139-42, 1999.
Article in English | MEDLINE | ID: mdl-10097700

ABSTRACT

OBJECTIVE: To study the cytologic findings of follicular variant of papillary thyroid carcinoma (FVPTC) and to compare them with the cytologic findings on other thyroid lesions. STUDY DESIGN: The study group consisted of aspirate smears from 15 cases of histologically proven FVPTC. The control group consisted of 152 cases, including adenomatous colloid goiter (70), usual papillary carcinoma (40), follicular adenoma (30), Hürthle cell neoplasm (7) and medullary carcinoma (5). RESULTS: The smears of FVPTC revealed numerous colloid balls in the background, multilayered microfollicles (rosettes), numerous nuclear grooves and inclusions in the monolayer sheets of follicular cells, very rare giant cells, absence of calcification and papillary clusters. Rosettelike microfollicles and numerous colloid balls were not seen in the control group. CONCLUSION: The combination of numerous colloid balls and rosettelike microfollicles was frequently seen in FVPTC. This combination was not observed in the control group.


Subject(s)
Carcinoma, Papillary, Follicular/diagnosis , Carcinoma, Papillary, Follicular/pathology , Thyroid Neoplasms/diagnosis , Thyroid Neoplasms/pathology , Adenoma/diagnosis , Adenoma/pathology , Adult , Biopsy, Needle , Diagnosis, Differential , Female , Goiter/diagnosis , Goiter/pathology , Humans , Inclusion Bodies/pathology , Male , Middle Aged
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