Analysis of the survival of patients with breast cancer depending on age, molecular subtype of tumor and metabolic syndrome.

AIM: To analyze the survival of patients with breast cancer (BC) depending on age, molecular subtype of the tumor and the presence of metabolic syndrome. PATIENTS AND METHODS: We have analyzed the results of examination and treatment of 202 patients with BC of stages I-III. The patients were distributed by age into 2 groups. The group 1 included 86 elderly patients (from 65 to 84 years old), the group 2 - 116 younger patients (from 32 to 64 years). An overall 1-, 3- and 5-year survival rates of the treated patients were assessed. The significance of factors influencing the overall survival (OS) of patients with BC was determined using the methods of statistical analysis. RESULTS: Molecular subtype of BC significantly affects survival rates: in a case of a luminal B subtype the 5-year OS was 71.6 vs 80% (p < 0.05) in groups 1 and 2, respectively while in a case of a basal-like subtype it was 60.2% and 71.6% (p < 0.05). The presence of metabolic syndrome significantly reduced the 5-year OS (up to 70.7% and 80.6%, p < 0.05 in groups 1 and 2, respectively). CONCLUSION: The OS is lower in elderly patients with BC compared with younger patients, especially in those who suffer from metabolic syndrome.

Molecular-genetic models for prognosis of development of tumors of reproductive system in women with family history of cancer.

AIM: To develop a prognostic molecular genetic model for assessing the risk of development of benign and malignant tumors of female reproductive organs (FRO) in patients from cancer-affected families. PATIENTS AND METHODS: The work presents the data on a comprehensive clinical examination of 210 women (90 patients with FRO cancer with aggregation of tumor pathology in families, 65 patients with benign pathology of FRO from cancer-affected families, 55 women - control group of healthy women without family history of cancer). Clinical genealogical analysis, morphological examination of tumors and molecular genetic studies of genomic DNA from peripheral blood and tumors were carried out. RESULTS: It was established that in the families of patients with benign and malignant pathology of FRO, malignant tumors associated with Lynch II syndrome are observed. Based on the analysis of detected ESR-1, CYP 2D6*4 and mutations in BRCA1/2 genes in cancer patients and in patients with benign pathology, molecular genetic models have been developed to assess the individual risk of development of benign and malignant tumors of FRO. It has been established that these molecular genetic models and combinations of gene mutations and gene polymorphisms (SNP) by the intergene interaction that was analyzed, were found to be reliable in assessing the risk of benign and malignant pathology of the mammary gland and ovary. CONCLUSIONS: The model, which included the polymorphic variants of the T397C(ESR1)/CYP 2D6*4 genes was of the best predictive accuracy for the evaluation of the risk of benign tumors of the FRO (71.68%) and the highest reliability (p < 0.001). At the same time, all identified models of intergene interaction in the development of malignant pathology of FRO were reliable, prognostically significant with high reproduction and almost identical accuracy (65.00-68.23%). The obtained results indicate a high informativeness of such molecular genetic indices as the polymorphism of ESR1 and CYP 2D6*4 genes and mutations in BRCA1/2 genes to assess the risk of benign or malignant tumors of FRO in families of patients with family history of cancer.

Association of CD44+CD24-/low with markers of aggressiveness and plasticity of cell lines and tumors of patients with breast cancer.

AIM: To search for additional molecular-biological markers of cancer stem cell (CSC) involved in the development of intra-tumor heterogeneity for the detection of features of the breast cancer (BC) pathogenesis. MATERIALS AND METHODS: Expression of estrogen receptors (ER), progesterone receptors (PR), Her2/neu, E- and N-cadherin, CD24, CD44, Bcl-2, Bax, Slug, P-gp, glutathione-S-transferase (GST) and metallothionein in cell lines was determined by the immunocytochemical method. Expression of ER, PR, Her2/neu, CD24 and CD44 in the surgical material of BC patients were determined by the immunohistochemical method. The levels of the miRNA were determined using real-time polymerase chain reaction. RESULTS: Cells of high-grade malignancy (HGM), MDA-MB-231 and MDA-MB-468 are characterized by high expression of stem cell markers compared to the cells of low-grade malignancy (LGM), T47D and MCF-7: CD44 levels in T47D and MCF-7 cells were in range of 72-79 points, which is significantly lower than in HGM cells (p < 0.05). Also, HGM cells with the properties of CSC were characterized by high expression of antiapoptotic proteins, the transcription factor Slug, and low levels of proapoptotic protein Bax (p < 0.05) compared to LGM cells. In cells with CSC characteristics an increased expression of transferrin and its receptor, ferritin, fentorin and hepcidin was revealed indicating activation of the endogenous iron metabolism. The characteristic feature of HGM cells with CSC phenotype were the increased levels of oncogenic miR-221, -155 and -10b by 60%, 92% and 78%, respectively, and decreased levels of oncosuppressive miR-29b, -34a and -200b by 8.4 ± 0.3, 4.6 ± 0.2, and 3.4 ± 0.6 times compared to MCF-7 line cells. It has been established that the development of resistance to cytostatics is accompanied by increased aggressiveness of tumor cells, loss of expression of hormonal receptors and acquiring of stem phenotype. In particular, increased expression of P-gp was observed in BC cells during the development of resistance to doxorubicin, of GST during the development of resistance to cisplatin along with increased CD44 expression (p < 0.05). We have revealed the relation between the presence of cells with the CSC phenotype (CD44+CD24-/low) and clinical and pathological characteristics of BC patients, their survival and BC sensitivity to neoadjuvant therapy (p > 0.05). CONCLUSIONS: The dependence between the expression of CSC markers and the degree of malignancy of tumor cells, development of resistance to cytostatics in vitro was established as well as the predictive value of the detection of the CSC for the individual prognosis of the BC course and sensitivity of the tumors to the treatment.

СD44+/CD24- markers of cancer stem cells in patients with breast cancer of different molecular subtypes.

AIM: To determine frequency of tumors with immunohistochemical markers of cancer stem cells (CSC) CD44+/CD24- in patients with breast cancer (BC) of different molecular subtype and to evaluate their prognostic value. OBJECT: Surgical material of 132 patients with BC stage I-II, age from 23 to 75 years, mean age - 50.2 ± 3.1 years was studied. METHODS: Clinical, immunohistochemical (expression CD44+/CD24-), morphological, statistical. RESULTS: BC is characterized by heterogeneity of molecular subtypes and expression of markers (CD44+/CD24-). Immunohistochemical study of expression of CSC markers in surgical material has detected their expression in 34 (25.4%) patients with BC of different molecular subtypes. The highest frequency of cells with expression of CSC marker was observed in patients with basal molecular subtype (44.8% patients). Most of BC patients with phenotype CD44+/CD24 had stage I of tumor process (34.3%). Statistical processing of data has showen that Yule colligation coefficient equaled 0.28 (р > 0.05) that argues poor correlation between stage of tumor process and number of tumors with positive expression of CSC markers. Statistical processing of data has showen high correlation between presence of cells with expression of CSC markers and metastases of BC in regional lymph nodes (Yule colligation coefficient equals 0.943; р < 0.5). Difference in overall survival of patients with BC of basal molecular subtype depending on expression of CSC CD44+/CD24- markers was detected. Survival of patients with basal BC was reliably higher at lack in tumors of cells with CSC markers CD44+/CD24- and, correspondingly, lower at presence of such cells (р < 0.05). In patients with BC of luminal (A and B), HER-2-positive subtypes, significant change in survival of patients depending on expression of CSC markers was not determined (р > 0.05). CONCLUSION: Significance of tumor cells with markers CD44+/CD24- within the limits of molecular subtype of BC may be additional criterion for advanced biological characteristic of BC, and in patients with BC of basal molecular subtype - for predictive evaluation of individual potential of tumor to aggressive clinical course.

Significance of adhesion molecules expression for estimation of serous ovarian cancer prognosis.

AIM: To study the adhesion molecules CD44 and E-cadherin expression in serous ovarian cancer (OC) and their relationship with clinicopathological peculiarities of tumor process and prognosis. MATERIALS AND METHODS: The study was performed on OC samples from 72 patients with serous OC of stages I-III. Expression of CD44 and E-cadherin in tumor samples was evaluated with the use of immunohistochemical analysis. RESULTS: Immunohistochemical analysis has revealed that in 58.3% of OC patients CD44 expression was detected in more than 10% of cells with average level of 30.0В±5.6%. E-cadherin expression was present in 47.2% of tumors, and 12.2В±3.6% cells were immunopositive. CD44 and E-cadherin expression depends on degree of cytomorphological malignancy and cell differentiation. A reverse correlation between CD44 and E-cadherin expression in primary serous OC (r=-0.38, p < 0.05) has been found. Increased CD44 expression (mean index) was not observed in peritoneal metastases compared to primary tumors, but analysis of individual indexes showed increase of CD44 expression in 46.6% OC patients with metastases. The dependence between overall survival of OC patients and the molecular phenotype of tumor cells, in particular, poor prognosis for OC patients with CD44(+)/E-cadherin(-) and CD44(+)/budding(+) tumor cells phenotype, has been revealed. CONCLUSION: The results of morphological and immunohistochemical analysis has shown the increase of adhesive features of serous ovarian cancer cells what may be significant for estimation of ovarian cancer aggressiveness and development of metastatic process.

[Morphologic and immunohistochemical features of Fallopian tubes and ovaries in women with genetic predisposition to ovarian cancer].

The fallopian tubes and ovaries samples from 5 breast cancer patients and 12 practically healthy women considered to be at high risk for developing ovarian cancer underwent evaluation for morphological features and immunohistochemical expression of Ki-67 and p53. In high-risk patients the multiple follicular, serous, epithelial inclusion cysts (often with epithelial hyperplasia), surface papillomatosis, fibromatosis, cortical stromal hyperplasia in ovaries and fibres' sclerosis, paratubal cysts in fallopian tubes were discovered. Immunohistochemical study revealed strong expression of Ki-67 (LI 16-53,6%) in epithelial cells of follicular and inclusion cysts in 25% (3/12) of practically healthy women of reproductive age. P53 expression was determined in epithelium of inclusion cysts with high proliferative activity in 16,6% (2/12) of practically healthy women with BRCAI/2 mutation. The current results suggest histological evidence for the existent of pre-neoplastic phenotype defined by above-described histologic features in the high-risk ovaries, especially in carriers of BRCA1/2 mutations. P53 and Ki-67 expression in epithelium of inclusion cysts in women with BRCA1/2 mutations indicates the early genetic alterations in these cells and high risk of malignant transformation.

[Cytogenetic changes in the Namalwa cell line of human malignant lymphoma induced by inhibitors of DNA replication and synthesis]. / Tsitogeneticheskie izmeneniia v kletkakh perevivaemoi linii Namalwa zlokachestvennoi limfomy cheloveka, indutsirovannye ingibitorami replikatsii i sinteza DNK.

Namalwa cells originating from the malignant human lymphoma have been analyzed cytogenetically upon short-time exposure to subtoxic doses of inhibitors of DNA replication and synthesis, either etoposide or fludarabine. The intact cells were characterized by the modal class of the chromosomes within the diploid range with the proportion of the aberrant cells amounting to 16.0 +/- 0.5%. Upon exposure to etoposide the percentage of the aberrant cells increased amounting to 26.1 +/- 2.9 through 39.8 +/- 1.7% depending on the duration of the exposure and the dose of the drug. At the same time the number of the polyploid cells increased but the modal class retained within the diploid range. Upon exposure to fludarabine the percentage of the cells with the aberrant chromosomes increased to 57.1 +/- 2.9%. Two modal classes appeared--the first approaching the diploid number and the second being polyploid. The exposure to either etoposide or fludarabine resulted in increasing number of the chromatide aberrations with more frequent involvement of #1, #2, #5, #6, #7, #11, #13, #14, #16 and #17 chromosomes. The data obtained have shown the susceptibility of Namalwa cells to the subtoxic concentrations of the inhibitors of DNA synthesis and replication used in the study resulting in the survival of the novel clones resistant to the drugs.

[Complex estimation of proliferative activity of epithelial cells of the large intestine damaged by polyps and cancer]. / Kompleksna otsinka proliferatyvnoï aktyvnosti klityn epiteliiu slyzovoï obolonky tovstoï kyshky vrazhenoï polipamy ta rakom.

Peculiarities of mitotic regime and expression of proliferating cell nuclear antigen were investigated in 18 polyps and 35 cases of colorectal cancer. Direct relationship between spectrum and degree of manifestation of proliferative activity, level of morphological malignant tumors and accumulation of oncopathology in the patient pedigrees was established.

[The determination of chromosome fragile sites in the peripheral blood lymphocytes of patients with colorectal cancer taking into account the cancer pathology predisposition in the familial anamnesis]. / Opredelenie khromosomnoi sait-lomkosti v limfotsitakh perifericheskoi krovi bol'nykh kolorektal'nym rakom s uchetom otiagoshchennosti semeinogo anamneza po onkopatologii.

Results of comparative study of spontaneous and 5-bromdeoxyuridine-induced fragility of peripheral blood lymphocytes chromosomes in 9 patients with colorectal adenocarcinoma were presented. It was shown the increase of average spontaneous level of chromosomal fragility in patients with tumor aggregation in family as well as without it to 4.5 +/- 1.0 and 5.3 +/- 1.1 per 100 tested cells, accordingly. The increase of average level of damaged chromosomes in spectrum of rare sites to 12.5 +/- 2.6 in the patients with tumor aggregation in pedigree comparing to the patients without oncopathology in family 8.0 +/- 1.7 was observed. The most number of rare fragile sites was observed in 1q21 site of the chromosome 1. Possible connection between fragile sites of chromosomes in normal cells and malignant processes in the patients with colorectal cancer is discussed.

[The characteristics of protein p53 expression in the mucosal epithelium of the large intestine affected by polyps and cancer]. / Osoblyvosti ekspresiï bilka p53 v epiteliï slyzovoï obolonky tovstoï kyshky, vrazhenoï polipamy ta rakom.

The direct dependence between p53 expression in epithelial cells and the level of morphological malignant neoplasm was established as the result of investigation of expression of this biomolecular marker in 12 polyps and 35 colorectal cancer. It was shown that family cancer syndrome 1 type was observed in pedigrees of majority patients with p53 positive cancers.

[The clinico-genetic characteristics of the origin and manifestation of colorectal cancer]. / Kliniko-henetychni osoblyvosti vynnyknennia ta manifestatsiï kolorektal'noho raku.

A clinical and genealogical investigation has been carried out in 169 subjects of both sexes, presenting with colorectal oncopathology. Particular features of the colorectal carcinoma in the patients have been recognized. The frequency of the tumours spread in the relatives of the probands is determined as is the spectrum of aggregation of large intestine carcinoma with neoplasia of other genesis in the pedigrees. The data obtained suggest an important contribution of genetic factors into the origination of colorectal carcinoma.

[The cytomorphological characteristics of the epithelial layer of the gastric and duodenal mucosa in those who worked in the cleanup of the aftermath of the accident at the Chernobyl Atomic Electric Power Station]. / Tsytomorfolohichna kharakterystyka epitelial'noho pokryvu slyzovoï obolonky shlunka ta dvanadtsiatypaloï kyshky u likvidatoriv naslidkiv avariï na ChAES.

The complex of cytomorphological changes (dystrophy, dysregeneration, dysplasia, metaplasia) and dysfunction of gastric and duodenal mucosa epithelium were found in 79 liquidators of the Chernobyl disaster. It was shown that the reparative capability of superficial epithelium was retained. The increased number of abnormal mitoses was observed. The highest proliferative activity of duodenal epithelium reflects the intensity of reparative processes in duodenum at acute phase of peptic ulcer disease.

[The functional characteristics of the chromosomal nucleolus organizer regions of patients with endometrial cancer]. / Osobennosti funktsionirovaniia iadryshkoobrazuiushchikh raionov khromosom u bol'nykh rakom éndometriia.

The nucleolus organizer region activity and the level of acrocentric chromosome association in cultured peripheral blood lymphocytes was studied in patients with endometrial glandular hyperplasia and endometrial cancer. The number of active nucleolus organizer regions and the frequency of 13 and 21 acrocentric chromosome associations was higher in patients with endometrial cancer.

[Structural chromosome aberrations in the peripheral blood lymphocytes in patients with precancer and endometrial cancer]. / Strukturnye aberratsii khromosom v limfotsitakh perifericheskoi krovi u bol'nykh predrakom i rakom éndometriia.

The level of chromosomal aberrations in cultured blood lymphocytes from patients with hyperplasia, atypical hyperplasia of endometrium and endometrial cancer was studied. Chromosomal aberration frequency significantly increased up to 5.000 +/- 00.47% in patients with atypical and endometrial hyperplasia and to 5.21 +/- 00.53% in patients with endometrial cancer in comparison to 0.67 +/- 0.21% in the control. All indices were more pronounced in endometrial cancer patients with pathology in pedigree.

[A cytogenetic examination of persons subjected to radiation exposure in certain regions of Ukraine]. / Tsitogeneticheskoe obsledovanie lits, podvergshikhsia radiatsionnomu vozdeistviiu v nekotorykh regionakh Ukrainy.

We have summarized the results of cytogenetic studies of peripheral blood lymphocytes conducted in the institutions of Kiev and Kharkov in persons irradiated after the accident of the Chernobyl NPP. The average level of chromosomal aberrations and the appearance of cytogenetic markers characteristic of their irradiation effect was higher in the examinees in comparison with the control. The increase in the number of aberrations in certain Kiev residents as well as the development of lymphogranulomatosis in some liquidators of the accident were detected. The increase in the frequency of chromosomal aberrations in liquidators of the accident was accompanied by the reduction in the level of reparative DNA synthesis of blood lymphocytes. The necessity to expand cytogenetic examination in persons that participated in the liquidation of the accident on the ChNPP and Kiev inhabitants is discussed.

[Polymorphism of the C-band segments of chromosomes 1, 9 and 16 in the peripheral blood lymphocytes of patients with endometrial cancer]. / Polimofizm C-segmentov 1, 9 i 16 khromosom v limfotsitakh perifericheskoi krovi bol'nykh rakom éndometriia.

The paper presents results of investigation of constitutive heterochromatin of chromosomes 1, 9 and 16 in 23 patients with endometrial cancer and 5 women of the control group. The analysis was carried out on slides obtained by the routine method of the peripheral blood cell culture. The C-band segment variability was studied by the C-binding method. The investigation established an increase of polymorphism of constitutive heterochromatin in cancer patients, extreme variants and heteromorphism of homologues of chromosomes 1, 9 and 16. It is shown that polymorphism of the C-band on chromosomes 1, 9 and 16 in patients with cancer in their families increases as compared to the patients without cancer pathology in pedigrees.

[Morphofunctional features of the nucleoli in glandular hyperplasia and endometrial cancer]. / Morfofunktsional'nye osobennosti iadryshek pri zhelezistoi giperplazii i rake éndometriia.

The morphofunctional peculiarities of epithelial cells nucleoli in patients with glandular hyperplasia and endometrial cancer has been studied on the cytological smears from endometrium stained with silver nitrate were investigated. The patterns of nucleolonemic nucleoli content and high activity of nucleolar organizers in cancer of corpus uteri are revealed.

[Familial-populational studies of endometrial cancer: segregation and genetic dispersion analyses]. / Semeino-populiatsionnye issledovaniia raka éndometriia: segregatsionnyi i genetiko-dispersionnyi analiz.

A clinico-genealogic investigation was carried out in 216 patients with endometrial cancer. Familial accumulation of endometrial and other cancer incidence was established. The segregation rates appeared to be lower than those expected from simple Mendelian models (2-11%). A multifactorial nature of endometrial cancer in overall susceptibility to the disease was found to be at 61%. A genetic correlation analysis showed endometrial cancer to share common genes with breast and gastric cancer in females. Tables of recurrent risk of the disease for relatives were prepared to be used in medico-genetic counseling.

[Cytogenetic changes in peripheral blood lymphocytes of oncology patients]. / Tsitogeneticheskie izmeneniia v limfotsitakh perifericheskoi krovi onkologicheskikh bol'nykh.

Analysis of home and foreign literature underlies the discussion of the significance of cytogenetic variations (frequency of aberrant cells and SCE-heteromorphism of C-chromatin and brittleness of chromosomes as indices of chromosome instability in oncological patients.

[Recurrent endometrial cancer after combined radiation therapy]. / Retsidiv raka éndometriia posle sochetannoi luchevoi terapii.

Data of a patients are reported who developed recurrence of highly differentiated cancer of the endometrium 20 years after combined. At that time the patient harboured also a borderline mucinous cystadenoma of the ovary with malignization foci. An analysis of the lymphocyte karyotype in the peripheral blood revealed a 14% increase of aberrant cells.