Исследование сывороточной микроРНК-122 при гепатите С и ассоциированной с ним гепатоцеллюлярной карциноме

BACKGROUND: The discovery of a cluster of short non-coding RNAs called microRNAs (miRNAs) has become an important event in molecular biology. One of its representatives, miR-122 plays a large role in regulating the expression of genes involved in carbohydrate, lipid metabolism, and iron metabolism in the body. In experimental studies it was shown that in addition to regulatory functions, miR-122 is involved in the pathogenesis of hepatitis C, providing the life cycle of the virus in the cell. The shift of emphasis in the study of miR-122 from basic research into clinical medicine seems to be a promising area of personalized medicine. AIMS: to determine the clinical significance of miR-122 in acute and chronic hepatitis C and associated hepatocellular carcinoma. MATERIALS AND METHODS: A total of 407 people were examined, including 17 patients with acute hepatitis C (AHC), 158 patients with chronic hepatitis C (CHC) and 62 patients with HCC associated with hepatitis C. Comparison groups consisted of 84 healthy individuals and 62 patients with clinically pronounced cirrhosis of a non-infectious etiology. In each cohort, the relative miR-122 level was determined in the blood of patients. The analysis was performed in PCR using the Qubit microRNA Assay Kit -100 for the quantitative determination of microRNAs (Thermo Fisher Scientific, USA). Relative miR-122 expression values were calculated by the formula 2 -CT using U6 snRNA as a reference RNA. RESULTS: The highest miR-122 level in serum was found in patients with AHC at the height of the icteric period. The level of miR-122 showed a direct correlation with the activity of hepatic transaminases in patients with AHC (r = 0.72) and CHC (r = 0.44). An analysis of miR-122 level relative to the degree of liver fibrosis in patients with chronic hepatitis C showed that, as liver fibrosis progresses, the level of miR-122 expression decreases. The decrease in miR-122 expression in patients with severe fibrosis was universal and did not depend on the etiology of the disease. The development of HCC in the presence of chronic hepatitis C was accompanied by a decrease in the level of miR-122 by 10 times on average compared to patients with chronic hepatitis C. CONCLUSIONS: The determination of the expression level of miR-122 in serum can be used in laboratory monitoring of the management of patients with HC as an indicator of the severity of liver damage in AHC and the rate of formation of liver fibrosis in CHC. Evaluation of possibility of using miR-122 as a predictor of the development of HCC in the outcome of HC requires additional studies of the specificity and sensitivity of the test and comparison of the obtained data with the results of using generally accepted protein tumor markers.Обоснование. Открытие кластера коротких некодирующих РНК, получивших название микроРНК (miRNAs), стало важным событием в молекулярной биологии. Один из его представителей ― miR-122 ― играет большую роль в регуляции экспрессии генов, задействованных в углеводном, липидном обмене и метаболизме железа в организме. В экспериментальных исследованиях показано, что помимо регуляторных функций miR-122 участвует в патогенезе гепатита С, обеспечивая жизненный цикл вируса в клетке. Смещение акцентов в изучении miR-122 с фундаментальных исследований в русло клинической медицины представляется перспективным направлением персонализированной медицины. Цель исследования ― определить клиническое значение miR-122 при остром и хроническом течении гепатита С и ассоциированной с ним гепатоцеллюлярной карциноме. Методы. Всего было обследовано 407 человек, в том числе 17 больных острым гепатитом С, 158 ― хроническим гепатитом С и 62 ― гепатоцеллюлярной карциномой, ассоциированной с гепатитом С. Группы сравнения составили 84 практически здоровых человека и 62 больных с клинически выраженным циррозом печени неинфекционной этиологии. В каждой когорте в крови больных было проведено определение относительного уровня miR-122. Анализ осуществляли методом полимеразной цепной реакции с использованием набора реактивов Qubit microRNA Assay Kit-100 для количественного определения микроРНК (Thermo Fisher Scientific, США). Относительные значения экспрессии miR-122 были рассчитаны по формуле 2 -CT с использованием U6 snRNA в качестве референсной РНК. Результаты. Наиболее высокий уровень miR-122 в плазме крови был обнаружен у больных острым гепатитом С в разгар желтушного периода. Уровень miR-122 в крови коррелировал с активностью печеночных трансаминаз у больных острым (r = 0,72) и хроническим (r = 0,44) гепатитом С. Анализ уровня miR-122 относительно степени фиброза печени у больных хроническим гепатитом С показал, что по мере прогрессирования фиброза печени снижается уровень экспрессии miR-122. Снижение экспрессии miR-122 у больных с выраженным фиброзом ― циррозом печени ― носило универсальный характер и не зависело от этиологии заболевания. Развитие гепатоцеллюлярной карциномы сопровождалось падением уровня miR-122 в среднем в 10 раз по сравнению с группой больных хроническим гепатитом С. Заключение. Определение уровня экспрессии miR-122 в крови может использоваться в лабораторном мониторинге ведения больных гепатитом С как показатель тяжести поражения печени при остром течении и скорости прогрессирования фиброза печени при хронических формах. Оценка возможности использования miR-122 в качестве предиктора развития гепатоцеллюлярной карциномы в исходе гепатита С требует проведения дополнительных исследований специфичности и чувствительности теста и сопоставления полученных данных с известными онкомаркерами протеиновой природы

Biomarkers in Diagnosis and Prediction of Hepatocellular Carcinoma Recurrence (Review)

International audienceHepatocellular carcinoma (HCC) is the second leading cause of death in oncological patients. The prognosis of the disease outcome depends directly on its timely detection. Currently, in the majority of countries, the diagnostic algorithm at the preclinical stage of tumor development includes determination of alpha-fetoprotein in combination with instrumental imaging techniques. This approach allows the detection of about 65-80% of liver tumors at an early stage (A according to the BCLC classification), whereas at a very early stage (0 according to the BCLC classification) only 32-50% of cases, the result which cannot be considered satisfactory. In this regard, the search for effective biomarkers of hepatocellular carcinoma is an important challenge that faces the world healthcare. Advances in proteomics and genomics have led to the discovery of numerous promising markers which are now being clinically tested. Molecules of protein nature proposed as hepatocellular carcinoma tumor markers in different periods of time are described in this review. Comparative data on their effectiveness and specificity are also presented. The possibility of isolated or combined use of these biomarkers for risk assessment and early diagnosis of primary liver cancer is considered

Synergistic induction of lipid catabolism and anti-inflammatory lipids in white fat of dietary obese mice in response to calorie restriction and n-3 fatty acids

Calorie restriction is an essential component in the treatment of obesity and associated diseases. Long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) act as natural hypolipidaemics, reduce the risk of cardiovascular disease and could prevent the development of obesity and insulin resistance. We aimed to characterise the effectiveness and underlying mechanisms of the combination treatment with LC n-3 PUFA and 10% calorie restriction in the prevention of obesity and associated disorders in mice. Male mice (C57BL/6J) were habituated to a corn-oil-based high-fat diet (cHF) for 2 weeks and then randomly assigned to various dietary treatments for 5 weeks or 15 weeks: (1) cHF, ad libitum; (2) cHF with LC n-3 PUFA concentrate replacing 15% (wt/wt) of dietary lipids (cHF + F), ad libitum; (3) cHF with calorie restriction (CR; cHF + CR); and (4) cHF + F + CR. Mice fed a chow diet were also studied. We show that white adipose tissue plays an active role in the amelioration of obesity and the improvement of glucose homeostasis by combining LC n-3 PUFA intake and calorie restriction in cHF-fed mice. Specifically in the epididymal fat in the abdomen, but not in other fat depots, synergistic induction of mitochondrial oxidative capacity and lipid catabolism was observed, resulting in increased oxidation of metabolic fuels in the absence of mitochondrial uncoupling, while low-grade inflammation was suppressed, reflecting changes in tissue levels of anti-inflammatory lipid mediators, namely 15-deoxy-Delta(12,15)-prostaglandin J(2) and protectin D1. White adipose tissue metabolism linked to its inflammatory status in obesity could be modulated by combination treatment using calorie restriction and dietary LC n-3 PUFA to improve therapeutic strategies for metabolic syndrome