BACKGROUND: Determination of the cytogenetic origin and morphological type of metastatic or primary tumor determines the prescription of therapy and affects the efficiency of patient’s treatment. Currently, a number of publications evaluate the potential of cytological diagnostics of melanoma. AIM: To evaluate the significance of diagnosing metastatic melanoma by fine-needle aspiration biopsy and primary melanoma by using the skin and mucosa imprints. METHODS: In a retrospective study, a comparative analysis of cytological melanoma samples, in comparison with clinical and anamnestic information and the results of histological, immunohistochemical, and molecular genetic studies, was carried out. Information about 109 patients from the cancer registry of the Altai Regional Oncology Dispensary (Barnaul, Russia) for 2022 was used in the study. Traditional and liquid-based methods for preparing samples were used. The samples were stained using Pappenheim and Papanicolaou methods. In some observations, cytological material was used for molecular genetic studies. Using the cancer registry data of the dispensary, the results of histological and of molecular genetic studies, and a final conclusion was given about each patient. RESULTS: Fine-needle aspiration biopsy was carried out in 80 patients. Tumor smears were obtained from 29 patients. The cytological diagnose “melanoma” was consistent with the data of histological and immunohistochemical studies (p 0.001) for all 109 patients. Melanoma was diagnosed for the first time in 66 (60.5%) patients. In other cases, the process of progression was noted within the period from one year to 20 years. Epidermal melanoma was noted in 101 (92.7%) cases, including 9 patients with acral melanoma, and 2 cases with localization on the vulva. Melanomas of mucosa were found in 5 cases (4.5%): in the rectum and anal canal, vagina, and in 2 cases on the hard palate. Non-epidermal (uveal) melanomas metastases were diagnosed in liver by fine-needle aspiration biopsy in 3 patients (2.8%). Based on the cellular composition, epithelioid cell melanoma was determined in 81 (74.3%) patients, mixed cell in 14 (12.8%) cases, spindle cell in 9 (8.2%), pigmentless in 2 (1.8%) cases and nevoid melanoma in 1 (0.9%) case. The mutation status was determined in 96 patients (88.1%). Of these, in 8 patients it was determined using cytological material. In epidermal melanomas, mutations in codon 600 of exon 15 of the BRAF gene were found in 43 (44.8%) patients, including 2 cases of acral melanoma. Mutations V600K, V600E/Ec were found in one patient each. In mucousal melanomas: Q61R mutation was found in exon 3 of the NRAS gene (vaginal melanoma), G12C mutation was identified in exon 2 of the NRAS gene (anal canal melanoma). In uveal melanomas, the assessed mutations were absent (it is necessary to determine mutation is GNAQ11 and BAP1 genes). CONCLUSION: Cytological diagnosis of melanoma by fine-needle aspiration biopsies and imprints from the tumor mass is a highly informative method that allows diagnosing melanoma and verifying the tumor subtype. The obtained results indicate tumor heterogeneity and differences in mutational status depending on the location of melanomas. The molecular classification of melanoma is important when choosing individualized therapy.

BACKGROUND: The duration and accuracy of verification of origin and morphological variant of adenocarcinoma in the cervical smear affect the effectiveness of the treatment of patients. Currently, few publications on the possibility of clarifying the origin of adenocarcinoma in a cervical smear using the traditional cytological method are not sufficient for evidence-based conclusions. AIMS: To identify patterns of cytological indicators of origin of adenocarcinoma in the biomaterial from the cervix. MATERIALS AND METHODS: In an observational one-stage retrospective study, a comparative analysis of cytological tests of cervical smears with the conclusion adenocarcinoma was carried out with clinical and anamnestic information and the results of histological, immunohistochemical, molecular and genetic examinations. Information about 143 patients in the cancer registry of the Altai Regional Oncological Dispensary (Barnaul) for 2021 was used for analysis. Cytology preparations were prepared by the traditional method, as well as by the method of liquid cytology, staining was carried out by the Papanicolaou (Pap Test) and Pappenheim methods. Testing for human papillomavirus (HPV) was carried out by immunohistochemical method and polymerase chain reaction. PIK3CA and KRAS mutations were detected by polymerase chain reaction. RESULTS: The cytological conclusion adenocarcinoma was confirmed by histology for all 143 women. There were cytology features of the morphological variant of AC, as well as to suggest the primary organ of adenocarcinoma, subsequently verified by visualization methods, histological, immunohistological examinations of biopsy and/or surgical material. The adenocarcinoma elements in the cervical smear was accompanied by adenocarcinoma in the endometrium, cervix, ovary/fallopian tube, colon (68.5, 17.6, 9.8 and 2.8% of cases, respectively; p 0.001). Differentiation of endometrial adenocarcinoma varied. Clear cell adenocarcinoma had specific cellular features. Cellular characteristics of HPV-associated and HPV-unassociated endocervical carcinomas differed from each other. Invasion of intestinal adenocarcinoma into the uterus and cervix was characterized by complexes with a palisade arrangement of cells in cervical smears. CONCLUSION: Cytolodiagnostics of adenocarcinoma in smear from the cervix has prospects for further improvement in the verification of the organ origin of carcinoma. It is assumed that it is rational to assess associations of cytology with histo-, cytochemical, molecular and genetic characteristics of adenocarcinoma.

This study was undertaken to analyze the effectiveness of cytological diagnostics of endocervical adenocarcinomas. We compared conventional liquid-based cytology, histology, immunohistochemistry, and molecular testing. A total of 25 endocervical adenocarcinomas, including endocervical adenocarcinomas in situ, were diagnosed using cytological methods over a year. Liquid-based cytology ensured better detection of glandular differentiation signs than conventional cytology. After molecular testing for human papillomavirus (HPV), we performed retrospective analysis of cytological characteristics of all endocervical adenocarcinomas (n = 15).We identified specific cellular characteristics of HPV-associated typical and mucinous adenocarcinomas. We also observed 1 case of non-HPV-related clear-cell and 1 case of non-HPV-related mesonephral adenocarcinoma.Our findings suggest that endocervical adenocarcinomas are a heterogeneous group of tumors. Endocervical adenocarcinomas accounted for 10.7 % of all primary cervical carcinomas (n = 214). Eighty percent of all endocervical adenocarcinomas were HPV-related, whereas the remaining 20 % were HPV-negative. We found no cytological differences between invasive endocervical adenocarcinomas and adenocarcinomas in situ.Mutations detected in some of the patients are an important diagnostic criterion that specifies whether the tumor is rare.

Retrospective analysis of the usage of fine needle aspiration breast cytolology has been represented in the present work. The potentialities of cytological diagnostics according to Yokohama system with characteristics of C1–C5 categories were estimated. The results of cytological conclusions of 4778 patients with breast lesions who had been examined in the Altay oncological dispensary during the year were studied. Fine needle aspiration breast cytology was used of outpatients in 89.6 % cases. The largest number of patients with pathological changes in the breast was noted in category C2 with benign processes (75.7 % of all cases). Difficult cases for cytological study, where the method could not guarantee the accuracy of the diagnosis, belong to the C3 and C4 categories (1.9 % of all cases). The cytological conclusion recommended the compulsory usage of the core biopsy. Malignant tumors were identified in 853 (19.9 %) patients with an indication of the histological type of tumors.Thus, the cytological technique (as a part of Triple test) should be chosen for outpatients with breast diseases using the Yokohama writing system (C1–C5 categories) of fine needle aspiration cytology.

The purpose of this article is to evaluate the possibilities of cytology for lung neuroendocrine tumors (small‑cell lung carcinoma and carcinoids) diagnostics. Cytology specimens obtained by bronchoscopy (n = 112), biopsy of metastatic lymph nodes (n = 27) or from pleural effusion (n = 8) were collected within over 1 year from 147 patients and studied. Small-cell lung carcinoma was diagnosed in 143 patients, representing 23,9 % of all lung carcinomas. The proportion of carcinoid tumors was 2,7 % of all neuroendocrine tumors. Typical carcinoid was observed in three cases, and atypical carcinoid — in one case. Cytologic features most significant for cytological diagnosis of small‑cell lung carcinoma and carcinoids were identified (n = 11). Discriminant analysis demonstrated that the proportion of accurate cytological diagnosis of small‑cell lung carcinoma and carcinoids was 96,69 %. Cytology is a reliable method for neuroendocrine tumor diagnosis. Immunocytochemistry with neuroendocrine markers along with light microscopy should be used to differentiate small‑cell lung carcinoma metastases from other tumors and non‑malignant pathology in pleural effusion specimens.

The aim of the study is to estimate the potentiality of using immunocytochemical and molecular genetic techniques in diagnosis of patients with pleural effusion. Materials and methods. The results of a cytological study of 580 patients with pleural effusion examined in the Altai Regional Oncological Dispensary in 2019 were evaluated. Immunocytochemical and molecular genetic techniques were applied using cytological specimens prepared from pleural fluid. Epidermal growth factor receptor (EGFR) gene status was determined. Results. Non-tumor pleural effusion was diagnosed in 378 cases (65%). Tumorous cells of 181 (31%) patients were found in pleural effusion. It was very difficult to diagnose a pathological process by cell composition in 21 (4%) cases because of the inability to distinguish histiocytes and mesothelial cells from tumor cells using light microscopy. Immunocytochemical researches of 105 patients with pleural effusion were used for specification of metastases and in uncertain diagnostic cases. EGFR gene mutation for the diagnosed pulmonary adenocarcinoma had been determined in 31 cases for proper assignment of the target medicines. Dotty L858R mutation and deletions 19 exon were detected. Conclusions: Light microscopy allowed us to differentiate tumor and non-tumor plural effusion in 96% of cases. Immunocytochemical technique revealed the primary tumor site in 95% of cases of pleural effusions. Cytological specimens are full-advantaged material for molecular genetic technique. Mutations were found in 19% of cases.

The aim of the study is to estimate potentiality of the cytological diagnostics of different types of mucin producing carcinomas of the breast.Materials and methods. Cytological evidences of the investigation of 35 patients with mucinous mass in cytological specimens were studied. Different types of mucinous carcinomas which had been diagnosed by cytological method were identified retrospectively.Results. “Pure” mucous carcinomas were divided into two types: hypocellular (A) and hypercellular (B) (n = 9 (25.7 %) and 14 (40 %), respectively). Mixed carcinomas (mucous carcinoma with invasive no special type carcinoma) were noticed of 12 women (34.3 %). 24 cell characteristics of mucin producing carcinomas were identified. Some cytological characteristics were estimated due to multiple factor analysis and discriminant analysis.Conclusions. Investigation results indicated that cytological technique is a high-quality diagnostic method. The fraction of correct classification of mucous carcinoma types based on discriminant analysis was 92–99 %.The study protocol was approved by the biomedical ethics committee of The Altai State Medical University, Ministry of Health of Russia.

Aim. This study aimed to improve the diagnostics of acral melanoma and investigate tumor morphological features of the chondroid and osseous matrix. Materials and methods. The article presents a clinical case of metastatic acral melanoma with chondroid and osseous metaplasia in a 60-year-old patient. Cytological, histologic, immunohistochemical, and molecular genetic studies were performed. Results. A conglomerate of lymph nodes was noted in the inguinal region of the patient. Oxyphilic chondroid masses and tumor cells with morphological features of sarcoma were revealed using thin-needle aspiration biopsy. On physical examination, pink-colored subcutaneous neoplasm was discovered in the skin of the heel. Histological examination of the primary neoplasm and inguinal lymph node was performed. Melanoma with chondro-osseous metaplasia was diagnosed. Immunohistochemical examination of the tumor revealed some pronounced diffuse expressions of S-100 protein, HMB-45, and focal expression of Melan A in tumor cells. The sample was tested to detect BRAF mutation, and no mutations were found. Conclusion. Morphological diagnostics of acral melanoma with chondro-osseous metaplasia is characterized by a high risk of diagnostic error. Therefore, additional immunohistological, molecular, and genetic studies should be used in the diagnostics of acral melanoma with chondro-osseous metaplasia.

This article presents the outcomes of our own research of the detection of circulating cells in the peripheral blood of 48 patients with oncological diseases, using hemocytofiltration. The observed circulating cells (21 cases, 43.8%) have been represented by three variants: more often in the form of “bare” atypical nuclei (76.2% cases), also single conserved tumor cells (14.3%) and large shapeless single cells, probably of non-epithelial nature (9.5%). The difficulties of identifying the obtained cells in light microscopy and technical aspects of the usage of immunocytochemical techniques have been discussed.

The article shows the possibilities of the usage of cytological material for diagnostics of lung cancer at the example of work of an oncological outpatient clinic during one year. Light microscopy was used to study of cytological specimens of various material of 721 patients. The method has allowed to determine the diagnosis of lung cancer in majority of the observed cases with definition of histological type in 89%. When comparing the results of cytology with the final diagnosis the difference of determination of the tumor histological types is less than 1%. The immunocytochemical test is used at the examination of pleural fluid of 40 patients without determined primary locus. The immunocytochemical research has allowed specifying the inhering of the tumor cells to the metastasis from the lung, elevating the precision of the diagnostics to 96%. The cytological material of 62 patients obtained during the bronchoscopic examination, puncture of lymphatic nodules, pleural fluid and sputum with presence of a sufficient quantity of tumor cells (not less than 200) represents appropriate material for molecular-genetic research. The usage of cytological material for search of somatic mutations is justified for oncological patients with locally advanced or disseminated process, whose cytological material is the unique accessible morphological material for research.

This article presents results of the introduction in practical oncology of molecular genetic investigations performed with the use of tumor DNA cells taken from the cytological specimens. There was investigated the molecular genetic characteristics of cytological specimens from 126 patients. In 80 cases with the proved diagnosis of pulmonary adenocarcinoma (n = 80) EGFR gene mutations were noted in 11.7% cases. KRAS, BRAF and BRCA1/2 gene mutations were determined in 46 women suffering from serous ovarian carcinoma. KRAS gene mutations in cells of ovarian low-grade serous carcinoma were determined in 62.5% of patients, BRAF- in 12.5% cases. BRCA1 gene mutations have been determined in 14.3% cases from the ovarian high-grade serous carcinoma group. In conditions of the presence of the sufficient amount of tumor cells the cytological material is the fully-featured material for molecular genetic investigations. The investigation both of EGFR gene mutations in pulmonary adenocarcinoma cases and KRAS, BRAF, BRCA1/2 gene mutations with serous ovarian carcinoma are mandatory in the appointment of targeted therapy.

23 © Коллектив авторов, 2017 *e-mail: cytolakod@rambler.ru В настоящее время молекулярно-генетические методы все шире используются для диагностики, прогноза и выбора оптимальной терапии при онкологических заболеваниях. Успехи молекулярной онкологии привели к созданию таргетных препаратов, воздействующих на внутриклеточные молекулярные мишени опухолевой клетки. Наиболее демонстративны успехи таргетной терапии при немелкоклеточном раке легкого, колоректальном раке, гастроинтестинальных стромальных опухолях, раке молочной железы, меланоме и других опухолях [4]. Одним из объектов, наиболее изученных в последние годы в качестве противоопухолевой мишени, является рецептор эпидермального фактора роста (EGFR). Открытие соматических мутаций гена EGFR явилось ключевым моментом в разработке стратегии лечения немелкоклеточного рака легкого и привело к появлению нового молекулярного показателя чувствительности опухоли легкого к ингибиторам тирозинкиназ [17, 19]. В 2004 г. в научной литературе появились сообщения об активирующих соматических мутациях в рецепторе эпидермального фактора роста. Исследователи обнаружили, что практически у всех пациентов, получающих положительный эффект от лечения гефитинибом и эрлотинибом, обнаружены мутации гена EGFR в ткани опухоли [16, doi: 10.17116/labs20176123-28

The results of diagnostics of 65 patients with serous ovarian carcinoma are given in the article. Analysis of the investigation has allowed to include 13 patients (20 %) in the group 1 (low grade) and 52 (80 %) in the group 2 (high grade). According to the results of the executed work it is determined that the Immunocytochemical technique allows to verify ovarian carcinoma, differentiating it with carcinoma metastases of the other organs, that is very important for following treatment. In case of sufficient tumour cells in cytological specimens, specimens are valuable material for molecular-genetic researches. According to the results of genetic DNA investigations, suitable for the estimation, there was absence mutation of V600E in the BRAF-gene in all cases. The genetic mutation of KRAS-gene with 5 patients from the group1 (low grade) was noticed. Among them the genetic mutation of G12V was detected in 4 cases and mutation of G12D in 1 case. The mutations in the BRCA1-gene were detected in 4 cases from the 2th group (high grade) three of them were presented by 5382insC mutation and one of them by T300G mutation. The usage of the multivariate analysis evaluating the cell composition of low and high grade carcinomas was allowed to distinguish the most significant features off ovarian carcinoma cells that will be allow to differentiate them by means of light microscopy.

The results of cytological diagnostics of 138 patients with adenocarcinoma of the uterine cervix were analyzed. Cytological verification of adenocarcinoma in specimens by the uterine cervix material allowed diagnosing as primary adenocarcinoma of the uterine cervix as well as metastatic lesions. The diagnose adenocarcinoma by the uterine cervix material was occurred in cases of 1) endometrioid carcinoma of 87 (63.0 %) patients; 2) endocervical carcinoma of 21 (15.2 %) patients; 3) metastases of serous ovarian carcinoma in 18 (13.0 %) cases; 4) other metastases-colon carcinoma of 6 (4.4 %) patients, rectal carcinoma 2 (1.4 %), glandular carcinoma including gastric signet ring cell carcinoma 3 (2.3 %), and mucinous carcinoma of unknown organs of 1 (0.7 %) patient. Particular features of the uterine cervix adenocarcinoma, endometrioid carcinoma and ovarian carcinoma due to data of discriminate analysis were not noticed. Metastatic intestinal adenocarcinoma has specific palisade-like cells located in complexes that allowed to verify carcinoma. Also, presence signet ring tumour cells allowed to diagnose the metastasis of gastric signet ring cell carcinoma.

The results of diagnostics of 65 patients with serous ovarian carcinoma are given in the article. Analysis of the investigation has allowed to include 13 patients (20 %) in the group 1 (low grade) and 52 (80 %) in the group 2 (high grade). According to the results of the executed work it is determined that the Immunocytochemical technique allows to verify ovarian carcinoma, differentiating it with carcinoma metastases of the other organs, that is very important for following treatment. In case of sufficient tumour cells in cytological specimens, specimens are valuable material for molecular-genetic researches. According to the results of genetic DNA investigations, suitable for the estimation, there was absence mutation of V600E in the BRAF-gene in all cases. The genetic mutation of KRAS-gene with 5 patients from the group1 (low grade) was noticed. Among them the genetic mutation of G12V was detected in 4 cases and mutation of G12D in 1 case. The mutations in the BRCA1-gene weredetected in 4 cases from the 2th group (high grade) three of them were presented by 5382insC mutation and one of them by T300G mutation. The usage of the multivariate analysis evaluating the cell composition of low and high grade carcinomas was allowed to distinguish the most significant features off ovarian carcinoma cells that will be allow to differentiate them by means of light microscopy.

The results of cytological diagnostics of 138 patients with adenocarcinoma of the uterine cervix were analyzed. Cytological verification of adenocarcinoma in specimens by the uterine cervix material allowed diagnosing as primary adenocarcinoma of the uterine cervix as well as metastatic lesions. The diagnose adenocarcinoma by the uterine cervix material was occurred in cases of 1) endometrioid carcinoma of 87 (63.0 %) patients; 2) endocervical carcinoma of 21 (15.2 %) patients; 3) metastases of serous ovarian carcinoma in 18 (13.0 %) cases; 4) other metastases-colon carcinoma of 6 (4.4 %) patients, rectal carcinoma 2 (1.4 %), glandular carcinoma including gastric signet ring cell carcinoma 3 (2.3 %), and mucinous carcinoma of unknown organs of 1 (0.7 %) patient. Particular features of the uterine cervix adenocarcinoma, endometrioid carcinoma and ovarian carcinoma due to data of discriminate analysis were not noticed. Metastatic intestinal adenocarcinoma has specific palisade-like cells located in complexes that allowed to verify carcinoma. Also, presence signet ring tumour cells allowed to diagnose the metastasis of gastric signet ring cell carcinoma.

The cytological characteristics of neoplastic pleurisies were studied in 119 patients with breast cancer. There were some cytological fea- tures of pleural fluid in breast cancer and particularities of the cell composition of pleurisies with ductal and lobular breast carcinoma. Differential diagnosis of epithelioid mesothelioma (1.7%) and ovary carcinoma (2.5%) was made and some difficulties were run with lobular carcinoma.