INTRATUMORAL AMPLIFICATION HETEROGENEITY IN HER2/neu-POSITIVE BREAST CANCER MOLECULAR-GENETIC SUBTYPES

The defining feature of HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtype breast cancer is HER2/neu gene amplification and protein overexpression on cancer cell membrane. The HER2-targeted therapy is nowadays available for patients with HER2-positive breast cancer However, a significant fraction of HER2+ tumors acquire or possess intrinsic mechanisms of resistance, based on multiple factors, and genetic heterogeneity among them. The aim of our study was to quantify the heterogeneity of HER2/neu amplification in HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtypes of breast cancer. Material and methods. A retrospective analysis of 210 cases referred for dual probe fluorescence in situ hybridization (FISH) confirmation of an immunohistochemical equivocal 2+ result was performed. Results. Our results demonstrated a heterogeneous amplification pattern of HER2/neu gene, whose expression is a substantial cause of HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtypes of breast cancer, in 31 % of invasive breast cancer cases. As heterogeneous, we interpreted tumors containing cells with HER2/CEP17 ratio < 2 and gene copies 4 ≤ HER2/neu < 6, that is, those without HER2/neu amplification. The amount of heterogeneous tumors between HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtypes was not statistically significant. ROC analyses identified optimal cutoff point for HER2/CEP17 ratio as 2.6 for distinguishing heterogeneous tumors. Conclusion. The heterogeneity of HER2/neu amplification is determined by FISH in 31 % of cases and is independent of molecular breast cancer subtype. If a HER2/neu-positive breast cancer has HER2/CEP17 ratio ≤ 2,6, it contains minor subclones without HER2/neu amplification with a probability of 95 %. Our results demonstrated that HER2/neu amplification heterogeneity may be important for prognosis of survival and treatment decisions

Значимость экспрессии онкопротеина Her-2/neu и компонента межклеточного матрикса тенасцина для метастатической активности рака мочевого пузыря

The authors conducted an immunohistochemical study with antibodies to Her-2/neu and tenascin in urothelial bladder carcinoma. The samples taken from 32 patients with stages Т2b/T3a, N0 and N+ urinary bladder carcinoma were studied. The tumor cells showed hyper-expression of the oncoprotein Her-2/neu in 6 out of 13 patients with metastases to the regional lymph nodes (T3b и T3a) and in 2 out of 7 tumors (Т3а) without lymph node involvement. Quantitative assessment of the reaction with tenascin antibodies revealed that in the metastatic lymph node involvement group, the staining area was much larger than that in metastasis-free group and it averaged 49.83 and 44.21% and 11.27 and 15.7%, respectively. Comparison analysis of Her-2/neu and tenascin expression revealed no clinically significant regularities. The study of the intercellular matrix showed a high correlation between the increase in the share of tenascin-expressing and/or containing structures and the rise in the metastatic activity of a tumor.

Современный подход к диагностике и оценке лечебного эффекта неоадъювантной терапии при раке молочной железы

Complete pathological tumor response is now considered the main criterion of effectiveness of neoadjuvant therapy and has great prognostic value. The paper describes the modern approach to the definition of residual tumor in breast cancer patients afterneoadjuvant chemotherapy, the criteria of a full morphological regression according to the latest clinical guidelines.Одним из основных методов оценки эффективности неоадъювантной терапии сегодня считается анализ полного патоморфологического ответа опухоли. Этот показатель имеет важное прогностическое значение. В работе рассмотрен современный подход к определению остаточной опухолевой нагрузки рака молочной железы после неоадъювантной химиотерапии, приведены критерии полного морфологического регресса согласно последним клиническим рекомендациям

Выявление ДНК вируса папилломы человека в поверхностной уротелиальной карциноме мочевого пузыря

An immunohistochemical research with antibodies to the mutant p53 protein and a revelation of the human papilloma virus DNA 16 and 18 types via in situ hybridization at the histological sections of the urothelial carcinoma are realized. The material of 44 patients with the superficial bladder cancer (Ta and T1 stages) and with a presence of the indirect signs of the viral infection was studied. 16 patients were included in the group of high risk recurrence, 13 patients were included in the group of the mean risk, 15 patients were included in the group of the low risk. HPV DNA was revealed in 12 of 44 cases only in patients of the mean and high risk groups (5 and 7 cases, respectively); all the positive results in the high risk group were with the HPV 16 type probe. Evaluation of the p53 protein showed a significant increase of its expression in the mean and high risk groups. p53 protein expression mean value was 23,67% in the low risk group, 36,53% in the mean risk group, 53,43% in the high risk group. Presence of HPV DNA was associated with the high p53 expression in the vast majority of cases.

Факторы апоптоза и пролиферации при раке почки

The problem of forecasting of a kidney cancer and a choice of a method of adjuvant treatment the advanced forms of this disease remains actual. In the given research the estimation of prognostic importance of apoptotic and proliferative markers in 66 patients with local advanсed and metastatic renal cell carcinoma has been led. Results of the work allow to conclude that the estimation of expression of these markers allows to allocate groups of patients of the high risk requiring careful supervision and treatment.

Up-regulation of expression and lack of 5' CpG island hypermethylation of p16 INK4a in HPV-positive cervical carcinomas

BACKGROUND: High risk type human papilloma viruses (HR-HPV) induce carcinomas of the uterine cervix by expressing viral oncogenes E6 and E7. Oncogene E7 of HR-HPV disrupts the pRb/E2F interaction, which negatively regulates the S phase entry. Expression of tumor suppressor p16(ink4a )drastically increases in majority of HR-HPV associated carcinomas due to removal of pRb repression. The p16(ink4a )overexpression is an indicator of an aberrant expression of viral oncogenes and may serve as a marker for early diagnostic of cervical cancer. On the other hand, in 25–57% of cervical carcinomas hypermethylation of the p16 INK4a promoter has been demonstrated using a methylation-specific PCR, MSP. To evaluate a potential usage of the p16 INK4a 5' CpG island hypermethylation as an indicator of tumor cell along with p16(ink4a )overexpression, we analyzed the methylation status of p16 INK4a in cervical carcinomas METHODS: Methylation status of p16 INK4a was analyzed by MSP and by bisulfite-modified DNA sequencing. The expression of p16(ink4a )was analyzed by RT-PCR and by immunohistochemical technique. RESULTS: The extensive methylation within p16 INK4a 5' CpG island was not detected either in 13 primary cervical carcinomas or in 5 cancer cell lines by bisulfite-modified DNA sequencing (including those that were positive by MSP in our hands). The number and distribution of rare partially methylated CpG sites did not differ considerably in tumors and adjacent normal tissues. The levels of the p16 INK4a mRNA were increased in carcinomas compared to the normal tissues independently of the number of partially methylated CpGs within 5'CpG island. The transcriptional activation of p16 INK4a was accompanied by p16(ink4a )cytoplasmic immunoreactivity in the majority of tumor cells and presence of a varied number of the p16 positive nuclei in different tumors. CONCLUSION: Hypermethylaion of the p16INK4a 5' CpG island is not a frequent event in HR-HPV-positive cervical carcinomas and cannot be an effective marker of cancer cells with up-regulated expression of p16(ink4a). Our data confirm other previous studies claiming specific p16INK4a up-regulation in the majority of cervical carcinomas at both the protein and mRNA levels. Cytoplasmic accumulation of p16(ink4a )is a feature of cervical carcinomas

ВИРУС ПАПИЛЛОМЫ ЧЕЛОВЕКА КАК ФАКТОР РИСКА ПРИ РАКЕ МОЧЕВОГО ПУЗЫРЯ

The paper reviews the data available in the literature on the possible involvement of human papillomavirus in the induction of bladder cancer (BC). The results of the authors studies in this area are summarized. The data obtained in clinical and experimental studies in vivo and in vitro count in favor of the idea that human papillomavirus may be implicated in the genesis of BC.Представлен обзор литературы по проблеме возможного участия вируса папилломы человека в индукции рака мочевого пузыря (РМП). Суммированы результаты собственных исследований в этой области. Данные, полученные на клиническом материале и экспериментальных моделях in vivo и in vitro, свидетельствуют в пользу точки зрения, согласно которой вирус папилломы человека может участвовать в генезе РМП

Оценка экспрессии PD-L1 у пациентов с уротелиальным раком, имеющих противопоказания к назначению препаратов платины

Background. Urothelial cancer ranks 7th and 17th of all the malignant tumors in males and females, respectively. Development of new immunotherapeutic drugs provides new possibilities in treatment of such patients, especially the patient population in whom platinum‑based therapy is contraindicated. Administration of immunooncology drugs requires determination of PD-L1, for which various diagnostic systems are used. The question regarding correlation of results of determination of PD-L1 expression remains of concern.Materials and Methods. The study was performed on 100 samples of surgical and biopsy samples of urothelial cancer. Two clones of 22C3 and SP142 with corresponding detection systems were used for the study. PD-L1 expression was assessed in tumor and immune cells.Results. The study demonstrated a high correlation of negative PD-L1 tumor status determined using both diagnostic agents (92 % and 97 %) and low correlation of results of positive PD-L1 status (67 % and 43 %).Conclusions. Thus, if a negative result of PD-L1 status of urothelial cancer is obtained using any of the diagnostic agents studied, repeated test with the other antibody is not required. If positive status is obtained in one test, the patient may have a negative status in the other test, which allows recommending a repeated testing in borderline cases using a test, recommended for the medicinal product untended for treatment.Уротелиальный рак мочевого пузыря во всем мире находится на 7 и Введение. Уротелиальный рак во всем мире находится на 7 и 17 местах среди всех злокачественных опухолей у мужчин и женщин соответственно. Появление новых иммунотерапевтических препаратов открывает новые возможности для лечения таких пациентов, особенно учитывая группу пациентов, имеющих противопоказания к назначению препаратов платины. Применение иммуноонкологических препаратов требует определения экспрессии PD-L1, для которого используются различные диагностические системы. Актуальным является вопрос о сопоставлении результатов определения экспрессии PD-L1 разными методами.Материалы и методы. Исследования проведено на 100 образцах операционного и биопсийного материала уротелиального рака. Для исследования были использованы два клона антител 22C3 и SP142 с соответствующими системами детекции. Оценивалась экспрессия PD-L1 в опухолевых и иммунных клетках.Результаты. Исследование показало высокое совпадение негативного PD-L1 статуса опухоли, выявленного при использовании обоих диагностикумов (92% и 97%) и низкое совпадение результатов по позитивному PD-L1 статусу (67% и 43%).Выводы. Таким образом, при получении негативного результата PD-L1 статуса уротелиального рака любым из исследованных диагностикумов не требуется повторное тестирование с другим антителом. При обнаружении позитивного статуса по одному тесту пациент может иметь негативный статус по другому тесту, что позволяет рекомендовать повторное тестирование в пограничных случаях с тестом, рекомендованным для предполагаемого для лечения препарата

Сравнение иммуногистохимических тестов в рамках исследования CLOVER Российского общества клинической онкологии

Introduction. The goal of the CLOVER study performed by the Russian Society of Clinical Oncology, was a pairwise comparison of three validated PD-L1 immunohistochemical (IHC) tests (Ventana SP142, Ventana SP263, Dako 22C3) in the patient population with non-small cell lung cancer (NSCLC). This study is the first large Russian comparative study to evaluate PD-L1 expression levels using immunohistochemistry methods.Materials and methods. The study was conducted on 473 NSCLC samples from Biobank. The IHC tests were carried out with 3 antibody clones. Four trained pathologists independently evaluated the percentage of positively stained tumor cells (TC) and immune cells (IC). To assess the correlation of TC and IC between different runs and the prognostic values of one test for another, a concordant analysis was used.Results. The number of PD-L1‑positive cells (≥1 %) was higher among IC compared with TC in all three IHC tests. Pearson correlation coefficients (PCC) for TCs were 0.71, 0.87, and 0.75 for 22C3 / SP142, 22C3 / SP263 and SP263 / SP142, respectively. PCC values for ICs were 0.45, 0.61, and 0.68 for the same pairs. A high coincidence of positive and negative results (>91 %) was obtained between the staining with antibodies 22C3 and SP263 of immunooncological agents in the 1st line.Conclusions. The highest correlation between IHC tests was obtained by pairwise comparison of 22C3 and SP263. Clone 22C3 can be considered as a substitute for SP263 in the first-line treatment of NSCLC. Clone SP142 showed weaker expression in TC and IC compared to the other two tests in patients with non-small cell lung cancer.Введение. Целью исследования CLOVER, проведенного Российским обществом клинической онкологии, было попарное сравнение трех валидированныхиммуногистохимических (ИГХ) тестов PD-L1 (Ventana SP142, Ventana SP263, Dako 22C3) на одной и той же популяции пациентов с немелкоклеточным раком легкого (НМРЛ). Данное исследование — это первое крупное росийское сравнительное исследование по оценке определения уровней экспрессии PD-L1 методами иммуногистохимии.Материалы и методы. Работа проведена на 473 образцах НМРЛ, полученных из Биобанка. Иммуногистохимическое исследование проведено с использованием 3 клонов антител. Четыре подготовленных патологоанатома независимо оценивали процентное содержание положительно окрашенных опухолевых клеток (ОК) и иммунных клеток (ИК). Для оценки корреляции ОК и ИК между различными анализами и прогностических свойств одного теста для другого был проведен конкордантный анализ.Результаты. Число PD-L1-позитивных клеток (1% и более) было выше среди ИК по сравнению с ОК во всех трех иммуногистохимических исследованиях. Коэффициенты корреляции Пирсона (PCC) для ОК составили 0,71, 0,87 и 0,75 между 22C3/SP142, 22C3/SP263 и SP263/SP142 соответственно. Значения PCC для ИК составили 0,45, 0,61 и 0,68 для тех же пар. Было получено высокое совпадение положительных и отрицательных результатов (>91%) между окрашиванием, полученным с антителами 22C3 и SP263 для иммуноонкологических препаратов в 1 линии.Выводы. Наиболее высокая корреляция ИГХ анализов была получена при попарном сравнении 22C3 и SP263. Клон 22C3 можно рассматривать в качестве замены SP263 при лечении НМРЛ в первой линии. Клон SP142 показал более слабую экспрессию в ОК и ИК по сравнению с двумя другими анализами у пациентов с плоскоклеточным раком

Implication of expression of the oncoprotein Her-2/neu and the intracellular matrix component tenascin for metastatic activity of urinary bladder cancer

The authors conducted an immunohistochemical study with antibodies to Her-2/neu and tenascin in urothelial bladder carcinoma. The samples taken from 32 patients with stages Т2b/T3a, N0 and N+ urinary bladder carcinoma were studied. The tumor cells showed hyper-expression of the oncoprotein Her-2/neu in 6 out of 13 patients with metastases to the regional lymph nodes (T3b и T3a) and in 2 out of 7 tumors (Т3а) without lymph node involvement. Quantitative assessment of the reaction with tenascin antibodies revealed that in the metastatic lymph node involvement group, the staining area was much larger than that in metastasis-free group and it averaged 49.83 and 44.21% and 11.27 and 15.7%, respectively. Comparison analysis of Her-2/neu and tenascin expression revealed no clinically significant regularities. The study of the intercellular matrix showed a high correlation between the increase in the share of tenascin-expressing and/or containing structures and the rise in the metastatic activity of a tumor