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Влияние индуцированного эпителиально-Ð¼ÐµÐ·ÐµÐ½Ñ Ð¸Ð¼Ð°Ð»ÑŒÐ½Ð¾Ð³Ð¾ Ð¿ÐµÑ€ÐµÑ Ð¾Ð´Ð° на экспрессию маркеров стволовости в ÐºÐ»ÐµÑ‚ÐºÐ°Ñ ÐºÐ¾Ð»Ð¾Ñ€ÐµÐºÑ‚Ð°Ð»ÑŒÐ½Ð¾Ð³Ð¾ рака

выпускная квалификационная работа бакалавра

Влияние индуцированного эпителиально-Ð¼ÐµÐ·ÐµÐ½Ñ Ð¸Ð¼Ð°Ð»ÑŒÐ½Ð¾Ð³Ð¾ Ð¿ÐµÑ€ÐµÑ Ð¾Ð´Ð° на экспрессию маркеров стволовости в ÐºÐ»ÐµÑ‚ÐºÐ°Ñ ÐºÐ¾Ð»Ð¾Ñ€ÐµÐºÑ‚Ð°Ð»ÑŒÐ½Ð¾Ð³Ð¾ рака

Abstract

Переход карцином в стадию метастазирования может быть осуществлен с помощью программы эпителиально-мезенхимального перехода (ЭМП). В данной работе было изучено влияние ЭМП на характеристики наиболее агрессивной популяции опухолевых клеток – опухолевых стволовых клеток (ОСК). Посредством введения плазмидной конструкции, содержащей клонированный ZEB1, был индуцирован ЭМП. Отбор стволового компонента осуществлялся с помощью репортерной конструкции, содержащей сайты связывания OCT4/SOX2. Методом ОТ-ПЦРв реальном времени были получены уровни мРНК основных маркеров ОСК. Уровни мРНК данных маркеров в клоне с индуцированным ЭМП были в несколько раз выше, чем в обычном стволовом компоненте. Несмотря на превышение уровней мРНК транскрипционных факторов TWIST и SNAIL, отвечающих за активацию ЭМП, в стволовом компоненте клеток не было выявлено отличий в уровнях мРНК ZEB1, виментина и Е-кадгерина, основных маркеров ЭМП. В то же время выявлены отличия в морфологии и миграционной способности клеток «подобных стволовым». Таким образом, можно заключить, что введение ZEB1 в клетки колоректального рака привело к получению клонов, не только характеризующихся повышенными уровнями маркеров ОСК, но и способных к миграции.

The epithelial-mesenchymal transition is the main program that is responsible for metastasis. The given work is devoted to the exploration of the induced epithelial-mesenchymal transition (EMT) on cancer stem-like cells (CSC) influence, which are the most aggressive members of the cancer cells population. We used a special plasmid, which contains the ZEB1 gene, to induce the EMT program in the colorectal cancer cell line MIP101. The population of cancer stem-like cells was selected using construction with six binding sites for OCT4/SOX2 transcription factors. We have used the real-time RT-PСR method to examine the main CSC marker’s mRNA levels. We noticed that mRNA levels of these markers in cancer stem-like cells with induced EMT was higher than in the control group of cancer stem-like cells. Also, we noticed that high mRNA levels of transcriptional factors such as TWIST and SNAIL, which are involved in EMT, had not affected the mRNA levels of ZEB1, vimentin and E-cadherin, which are the EMT markers. On the other hand, there were some differences in morphology and migration ability of cancer like-stem cells under EMT and control group. Thus, it can be concluded that ZEB1 transfection in colorectal cancer cells can lead to increased levels of CSC’s markers as well as high migration ability.

Keywords

cancer stem cells, tumorigenesis, колоректальный рак, transcription factors, транскрипционные факторы, epithelial-mesenchymal transition, эпителиально-Ð¼ÐµÐ·ÐµÐ½Ñ Ð¸Ð¼Ð°Ð»ÑŒÐ½Ñ‹Ð¹ Ð¿ÐµÑ€ÐµÑ Ð¾Ð´, colorectal cancer, Ð¾Ð¿ÑƒÑ Ð¾Ð»ÐµÐ²Ñ‹Ðµ стволовые клетки, канцерогенез

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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Average
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