oncotomskСибирский онкологический журналSiberian journal of oncology1814-48612312-3168Tomsk National Research Medical Сепtеr of the Russian Academy of Sciences10.21294/1814-4861-2022-21-1-47-56oncotomsk-2027Research ArticleЛАБОРАТОРНЫЕ И ЭКСПЕРИМЕНТАЛЬНЫЕ ИССЛЕДОВАНИЯLABORATORY AND EXPERIMENTAL STUDIESМутационный профиль КRAS-позитивного колоректального ракаMutational profile of KRAS-positive colorectal cancerhttps://orcid.org/0000-0002-0370-8667ТелышеваЕ. Н.TelyshevaE. N.

Телышева Екатерина Николаевна, кандидат биологических наук, патологоанатомическое отделение

SPIN-код: 8700-1335. Research ID: X-9043-2018. Author ID (Scopus): 57193142191

Россия, 125047, г. Москва, 4-я Тверская-Ямская ул., 16

Ekaterina N. Telysheva, PhD, Pathoanatomical Department

SPIN-соде: 8700-1335. Research ID: X-9043-2018. Author ID (Scopus): 57193142191

16, 4th Tverskaya-Yamskaya St., 125047, Moscow, Russia

https://orcid.org/0000-0002-7882-2579ШайхаевЕ. Г.ShaikhaevE. G.

Шайхаев Евгений Гаджирамазанович, кандидат биологических наук, патологоанатомическое отделение

SPIN-код: 5504-8523. Research ID: AAB-4981-2020. Author ID (Scopus): 55053528200

Россия, 125047, г. Москва, 4-я Тверская-Ямская ул., 16

Eugene G. Shaikhaev, PhD, Pathoanatomical Department

SPIN-code: 5504-8523. Research ID: AAB-4981-2020. Author ID (Scopus): 55053528200

16, 4th Tverskaya-Yamskaya St., 125047, Moscow, Russia

https://orcid.org/0000-0002-2584-802XСнигиреваГ. П.SnigirevaG. P.

Снигирева Галина Петровна, доктор биологических наук, патологоанатомическое отделение

SPIN-код: 4247-0600. Research ID: Y-4302-2018. Author ID (Scopus): 6602153865

Россия, 125047, г. Москва, 4-я Тверская-Ямская ул., 16

Galina P. Snigireva, DSc, Pathoanatomical Department

SPIN-code: 4247-0600. Research ID: Y-4302-2018. Author ID (Scopus): 6602153865

16, 4th Tverskaya-Yamskaya St., 125047, Moscow, Russia

G.Snigireva@nsi.ruФГАУ «Национальный медицинский исследовательский центр нейрохирургии им. академика Н.Н. Бурденко» Минздрава РоссииРоссияN.N. Burdenko National Medical Research Center of Neurosurgery of the Ministry of Health of the Russian FederationRussian Federation2022030320222114756Copyright © Телышева Е.Н., Шайхаев Е.Г., Снигирева Г.П., 20222022Телышева Е.Н., Шайхаев Е.Г., Снигирева Г.П.Telysheva E.N., Shaikhaev E.G., Snigireva G.P.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.siboncoj.ru/jour/article/view/2027

Цель исследования – изучение особенностей молекулярно-генетического профиля KRAS-позитивного колоректального рака (КРР).

Материал и методы

Материал и методы. В исследование было включено 42 пациента с диагнозом колоректальный рак, в опухолевой ткани которых методом ПЦР в режиме «реального времени» выявлена мутация в гене KRAS. С помощью технологии секвенирования нового поколения (NGS ) на платформе Illumina были проанализированы гены, участвующие в молекулярном патогенезе КРР: KRAS, BRAF, NRAS, APC, TP53, SMAD2, SMAD4, FBXW7, PIK3CA, CTNNB1, TCF7L2, MLH1, MSH2, MSH3, MSH6, ATM, TGF-BR2, AKT1, CDC27, CASP8, MAP2K4, DCC, DMD, MAP7, ERBB2, P3H3, MIER3, CADM1, FLT4, PTPN12, PIK3R1, EP300. Пробоподготовку библиотек из выделенной ДНК проводили с использованием коммерческих наборов GeneRead DNAS eq Targeted Panel v2 Human Colorectal Cancer («Qiagen», США); NEBNext Ultra DNA library Prep kit for Illumina и NEBNext Multiplex Oligos for Illumina («New England BioLabs»).

Результаты

Результаты. У 36 пациентов с KRAS-позитивным КРР зарегистрированы изменения в 13 генах, участвующих в молекулярном патогенезе заболевания. Всего было выявлено 82 соматические мутации. При этом у 9 больных дополнительно выявлено по одной мутации, у 17 – по 2 мутации, у 7 – по 3 мутации и у 3 – по 4 мутации. Сочетание сразу трех мутаций в ключевых генах, отвечающих за патогенез КРР (KRAS, APC и TP53), было выявлено у 15 (36 %) больных. Сочетание двух мутаций в генах KRAS и APC выявлено у 10 (24 %) больных, в генах KRAS и TP53 – у 8 (19 %) больных. Самое большое количество соматических мутаций выявлено в генах APC (59,5 %) и TP53 (54,7 %). Показано, что сочетание трех мутаций в ключевых генах (KRAS, APC и TP53) является наиболее неблагоприятным фактором прогноза и может свидетельствовать о более высокой агрессивности опухолевого процесса.

Заключение

Заключение. Полученная с помощью метода NGS информация о мутационном статусе KRAS-позитивной опухоли пациентов с КРР позволяет с учетом клинических характеристик персонифицировать тактику лечения, а также прогнозировать его течение.

Aim

Aim: to study the features of the molecular genetic profile of KRAS-positive colorectal cancer (CRC).

Material and Methods

Material and Methods. The study included 42 patients diagnosed with colorectal cancer. The KRAS gene mutation was detected in tumor tissue of these patients by real-time PC R. Using the next generation sequencing technology (NGS ) on the Illumina platform, the genes involved in the molecular pathogenesis of colorectal cancer, namely KRAS, BRAF, NRAS, APC, TP53, SMAD2, SMAD4, FBXW7, PIK3CA, CTNNB1, TCF7L2, MLH1, MSH2, MSH3, MSH6, ATM, TGF-BR2, AKT1, CDC27, CASP8, MAP2K4, DCC, DMD, MAP7, ERBB2, P3H3, MIER3, CADM1, FLT4, PTPN12, PIK3R1, and EP300 were analyzed. Sample preparation of libraries from isolated DNA was carried out using commercial kits GeneRead DNAS eq Targeted Panel v2 Human Colorectal Cancer (Qiagen, USA ); NEBNext Ultra DNA library Prep kit for Illumina and NEBNext Multiplex Oligos for Illumina (New England BioLabs).

Results

Results. In 36 patients with KRAS-positive tumors, changes were observed in 13 genes involved in the molecular pathogenesis of colorectal cancer. A total of 82 somatic variants were identified. Moreover, 9 patients additionally had one mutation each, 17 patients had 2 mutations each, 7 patients had 3 mutations each, and 3 patients had 4 mutations each. Combination of three mutations in key genes involved in the pathogenesis of colorectal cancer (KRAS, APC и TP53) was detected in 15 (36 %) patients. Combination of two mutations in the KRAS and APC genes was detected in 10 (23.8 %) patients, and in the KRAS and TP53 genes – in 8 (19.1 %) patients. The largest number of somatic mutations was found in the APC (59.5 %) and TP53 (54.7 %) genes. It was hown that a combination of three mutations in key genes was the most unfavorable prognosis factor and indicated a higher aggressiveness of the tumor process.

Conclusion

Conclusion. The information obtained using the NGS method on the mutational status of a KRAS -positive tumor in patients with colorectal cancer allows for personalized treatment as well as predicting the outcome.

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The authors declare that there are no conflicts of interest present.