e21067 Background: The purpose of the study was to analyze the expression of angiogenesis-controlling markers (CD31, VEGF) in tissues of melanocytic skin lesions. Methods: The study included 20 patients with benign pigmented lesions and 15 patients with superficial spreading and nodular melanoma. Immunohistochemical study was performed on sections of paraffin blocks for standard morphological studies using murine monoclonal CD31 and VEGF antibodies. UltraVision Quanto Detection System HRP DAB was used for visualization. Angiogenesis in tumors was assessed by intratumoral microvessel density (iMVD) index. Microvessels were stained with anti-CD31 antibody. The number of vessels was determined in each area of sight using objective lens with 40x magnification and AxioVs40 v-4.8.1.0 program. The tumor was considered VEGF-positive if more than 25% of tumor cells showed positive staining, and percentages of these tumor cells were calculated in each case (in %). Results: Quantitative evaluation of iMVD showed that the median number of microvessels in an area of sight in melanoma and nevi was 10.0±0.9 and 4.1±0.4, respectively, results of CD31 expression were statistically significant (р˂0.05). The numbers of CD31-stained vessels in melanoma ranged from 4 to 18 in one area of sight. 14 nevus cases (70%) showed 4 and less vessels, and only giant-cell nevi (6-30%) demonstrated up to 15 vessels in one area of sight. VEGF expression was found in the cytoplasm of melanoma cells in 100% of cases (15), in nevi – in 65% (13). The expression was not found in nevi cells in 35% (7). The number of VEGF-positive cells varied greatly in both melanomas and in nevi – from less than 5% to 50%. The maximal VEGF expression was observed in melanomas – 32.5±2.9%, while in nevi 23.8±2.1% (р˂0.05). Conclusions: The immunohistochemical study demonstrated a high angiogenic potential of melanoma that can be used as an additional criterion for tumor progression and malignant transformation of melanocytic skin neoplasms and thus contribute to the development of novel approaches to the diagnosis of the disease.
e23545 Background: The tumor ability to induce and maintain angiogenesis is one of the main stages of its development. Studies of molecular mechanisms of angiogenesis showed that the dynamic balance that ensures the formation and development of new vessels inside the tumor depends on pro- and anti-angiogenic factors. VEGF and the CD34 endothelial cell marker are considered among the main activators of angiogenesis. Our purpose was to study characteristics of angiogenesis in primary and recurrent soft tissue sarcomas. Methods: The study included 30 patients with primary sarcomas (group 1) and 26 patients with recurrent sarcomas (group 2). Sections of tissues embedded in paraffin blocks were studied by immunohistochemistry using the Thermo Scientific 480S automated stainer. The Reveal Polyvalent HRP-DAB Detection System was used for the visualization. The density of blood vessels stained with antibodies against VEGF and CD34 was determined. The statistical analysis of results was performed in the STATISTICA 10.0 program (StatSoftInc., USA). Results: The numbers of blood vessels ranged: CD34 in group 1 – 7-16 blood vessels in one field of view; in group 2 – 2-18 vessels. VEGF in group 1 – 2-20 vessels, with up to 40 vessels in 3 cases (11.5%) only; in group 2 – 5-11 vessels, with up to 20 vessels in 2 cases (7.7%). The average numbers of microcirculatory vessels stained with antibodies against VEGF and CD34 were: CD34 –12.2±1.04 and 8.4±1.4 (p = 0.0247) in groups 1 and 2, respectively; VEGF – 12.8±4.06 and 8.4±11.6 (p = 0.3074) in groups 1 and 2, respectively. Conclusions: The IHC analysis showed the minimal numbers of microcirculatory vessels in one field of view in patients with recurrent soft tissue sarcomas. The value was 1.5 times lower for both CD34 and VEGF, compared to patients with primary tumors. However, only CD34 value was statistically significant (the Mann–Whitney U-test). Probably, a certain decrease in angiogenesis factors in recurrent tumors can be explained by adjuvant chemotherapy suppressing tumor growth.
Ростовский научно-исследовательский онкологический институт, г. Ростов-на-Дону, Россия Реферат Цель. Исследовать зависимость количества циркулирующих опухолевых клеток в периферической крови у больных колоректальным раком от клинико-морфологических характеристик основного заболевания. Методы. В исследование был включён 91 пациент с верифицированным метастатическим колоректальным ра-ком Т3-4N1-2М1. Средний возраст больных составлял 61,5±1,7 года. Пациенты были разделены на основную группу (лапароскопическое хирургическое лечение, n=44) и контрольную (хирургическое вмешательство откры-тым доступом, n=47). Число циркулирующих опухолевых клеток определяли в системе CellSearch TM в перифе-рической крови, взятой до оперативного вмешательства. Изучение ассоциации признаков по методу построения таблиц сопряжённости заключалось в расчёте коэффициента взаимной сопряжённости Пирсона c 2 с поправкой Мантеля-Хэнзеля на правдоподобие (непараметрическая поправка), оценке статистической значимости сопря-жённости и анализе тесноты ассоциации по коэффициенту взаимной сопряжённости А. Чупрова. Результаты. Нами обнаружена сопряжённость числа циркулирующих опухолевых клеток с клинико-морфоло-гическими параметрами больных колоректальным раком. Выявлена взаимосвязь потенциальных факторов ри-ска и повышения в периферической крови количества циркулирующих опухолевых клеток для всех больных колоректальным раком, независимо от способа хирургического вмешательства. Показано, что наиболее выра-женная ассоциация числа циркулирующих опухолевых клеток в периферической крови до операции у больных с метастатическим колоректальным раком по коэффициенту взаимной сопряжённости (К) сложилась с наличи-ем отдалённых метастазов (статус M1b; К=0,63, р=0,0001) и стадией Т4 (К=0,56, р=0,0009). Вывод. Полученные результаты подчёркивают важную предикторную значимость уровня циркулирующих опухолевых клеток в периферической крови для оценки потенциала прогрессирования колоректального рака. Ключевые слова: циркулирующие опухолевые клетки, колоректальный рак, регионарные и отдалённые мета-стазы, степень дифференцировки. Circulating tumor cells and their relationship with clinical and morphological characteristics of colorectal cancer O.I. Kit, V.E. Kolesnikov, R.E. Tolmakh, I.A. Novikova, O.G. Shul'gina, E.F. Komarova, A.A. Demidova Rostov Research Institute of Oncology, Rostov-on-Don, RussiaAim. To investigate the dependence of the number of circulating tumor cells in peripheral blood of colorectal cancer patients on the clinical and morphological characteristics of underlying disease. Methods. 91 patients with verified metastatic colorectal cancer Т3-4N1-2М1 were included in the study. The average age of the patients was 61.5±1.7 years. The patients were divided into the study group (laparoscopic surgical treatment, n=44) and control group (open surgical intervention, n=47). The number of circulating tumor cells was determined in CellSearch™ system in the peripheral blood drawn before the intervention. The study of the association of attributes by construc...
e15531 Background: High incidence and mortality rates of gastric cancer cause a constant search for the most informative and effective methods of diagnosis and treatment assessment. In this regard, studying the expression of markers with the stem phenotype in primary tumor tissues in patients with gastric cancer with and without metastases is of undoubted interest. Methods: The study included 20 gastric cancer patients aged 30-80 years: group 1 – gastric cancer T3-4аN0-3M0 (G2) without metastasis (58.9±9.7 years); group 2 – gastric cancer T3-4аN0-3M1 (G2) with peritoneal metastasis (53.4±11.9 years). Immunohistochemical study was performed on paraffin-embedded tumor tissue sections using mouse monoclonal antibodies to CD44 (156-3С11 Thermo Scientific) at a 1:2500 dilution and rabbit polyclonal antibodies to CD133 (Cloud-Clone Corp.) at a 1:700 dilution; the Thermo Scientific autostainer was used for staining. Membrane staining and staining intensity were assessed: 0, 1+ weak, 2+ moderate, 3+ strong staining. Positive expression was defined as ≥10% cut-off for CD44 and ˃5% for CD133. Results: Positive expression of CD44+ was detected in 67% (13) in group 2 vs. 20% (4) in group 1. In the metastatic group, the number of cells that stained positive for CD44 expression ranged from 9 to 15%, on average 10.0±3.08%, without metastases – from single cells to 13%, on average 6.0±2.3%. A chi-square test showed statistically significant association in the groups (8.256 at p = 0.004). Positive CD133+ expression in tumor tissues was registered in 100% (20) in group 2 and 80% (16) in group 1. The range of positively stained cells in group 2 was from 10 to 40%, on average 21.3±11.6%, in group 1 - from single cells to 14%, on average 10.0±2.4%. A chi-square test showed statistically significant association in the groups (4.444 at p = 0.036). Conclusions: Immunohistochemical study of the selected tumor cell markers in gastric cancer revealed some characteristics of their expression depending on the presence of metastases. The results can be the basis for further research for the most complete characterization of a heterogeneous tumor population in gastric cancer and the role of individual cells in the tumor growth, progression and metastasis.
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