Анализ микроРНК miR-125a-5p, -27a-5p, -193a-5p, -135b-5p, -451а, -495-3р и -136-5р в клетках рака яичника и секретируемых ими экстраклеточных везикулах

Введение. Поиск маркеров для жидкостной диагностики рака яичника (РЯ) является одной из наиболее актуальных задач онкогинекологии. В настоящее время большой интерес в качестве источника онкомаркеров, в том числе микроРНК, вызывают экстраклеточные везикулы (ЭКВ). Ранее мы показали, что уровень miR-125a-5p, -27a-5p, -193a-5p и -135b-5p достоверно повышен, а miR-451а, -495-3р и -136-5р значимо снижен в ЭКВ маточных аспиратов больных РЯ.

Цель исследования – анализ уровней miR-125a-5p, -27a-5p, -193a-5p, -135b-5p, -451а, -495-3р и -136-5р в клеточных линиях РЯ и секретируемых ими ЭКВ.

Материалы и методы. Проведены культивирование клеточных линий РЯ (OVCAR-3, OVCAR-4, OVCAR-8 и SKOV3), выделение ЭКВ из кондиционированной среды методом ультрацентрифугирования, валидация ЭКВ с помощью анализа траекторий наночастиц (NTA), трансмиссионной электронной микроскопии и вестерн-блот-анализа экзосомальных маркеров. Также выполнены выделение микроРНК из клеток и ЭКВ, анализ микроРНК методом полимеразной цепной реакции с обратной транскрипцией в реальном времени в модификации Stem-loop.

Результаты. В клетках исследуемых линий РЯ экспрессия молекул miR-125a-5p, -27a-5p, -193a-5p и -135b-5p значительно превышала экспрессию miR-451а, -495-3р и -136-5р. Все линии клеток РЯ характеризуются соотношением «клетки >ЭКВ» для высоко экспрессируемых микроРНК и «ЭКВ >клетки» для низко экспрессируемых микроРНК.

Заключение. Результаты исследования свидетельствуют о связи дифференциальной экспрессии исследуемых микроРНК с патогенезом РЯ и подтверждают высокий диагностический потенциал данных молекул.

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