Kazan medical journalKazan medical journalКазанский медицинский журнал0368-48142587-9359Eco-Vector67860110.17816/KMJ678601MCWRCOExperimental medicineЭкспериментальная медицинаResearch ArticleMeta-analysis of transcriptome profiles of B16 melanoma cells after in vivo treatment with dacarbazineМетаанализ транскриптомных данных клеток меланомы B16 после воздействия дакарбазином in vivo体内暴露于达卡巴嗪后B16黑素瘤细胞转录组学数据的荟萃分析https://orcid.org/0000-0002-7226-95657656-8584LapkinaEkaterina Z.ЛапкинаЕкатерина Зиядхановна
Russian Federation

Assistant Professor, Depart. of Pathological Physiology named after prof. V.V. Ivanov

доцент, каф. патологической физиологии имени профессора В.В. Иванова

e.z.lapkina@mail.ruhttps://orcid.org/0000-0001-7085-63045810-5926ZinchenkoIvan S.ЗинченкоИван Сергеевич
Russian Federation

Senior Lab Assistant, Depart. of Pathological Physiology named after prof. V.V. Ivanov

старший лаборант, каф. патологической физиологии им. проф. В.В. Иванова

zinchenko.ivan.003@gmail.comhttps://orcid.org/0000-0003-3762-69746452-2085BondarEvgeniya I.БондарЕвгения Ивановна
Russian Federation

Cand. Sci. (Biology), Junior Research Associate, Lab. of Genomic Research and Biotechnology, Senior Lecturer, Depart. of Genomics and Bioinformatics

канд. биол. наук, младший научный сотрудник, лаб. геномных исследований и биотехнологии, старший преподаватель, каф. геномики и биоинформатики

bondar.zhenya.iv@gmail.comhttps://orcid.org/0000-0001-8142-42835412-2148RukshaTatiana G.РукшаТатьяна Геннадьевна
Russian Federation

MD, Dr. Sci. (Medicine), Professor, Head, Depart. of Pathological Physiology named after prof. V.V. Ivanov

д-р мед. наук, профессор, заведующая, каф. патологической физиологии им. проф. В.В. Иванова

tatyana_ruksha@mail.ruProfessor V.F. Voino-Yasenetsky Krasnoyarsk State Medical UniversityКрасноярский государственный медицинский университет им. проф. В.Ф. Войно-ЯсенецкогоSiberian Federal UniversityСибирский Федеральный университетKrasnoyarsk Science Centre of the Siberian Branch of Russian Academy of ScienceКрасноярский научный центр Сибирского отделения Российской академии наук130920250510202510657767860405202511062025Copyright ©; 2025, Eco-VectorCopyright ©; 2025, Эко-ВекторCopyright ©; 2025,2025Eco-VectorЭко-Векторhttps://kazanmedjournal.ru/kazanmedj/article/view/678601

BACKGROUND: Phenotypic plasticity and heterogeneity of melanoma cells arising from epigenetic regulation and the activity of transcription factors is pivotal in chemoresistance. Therefore, there it is necessary to explore the molecular mechanisms underlying resistance to alkylating agents, such as dacarbazine.

AIM: This study aimed to analyze transcriptome profiles of B16 melanoma cells after in vivo treatment with dacarbazine to identify key clusters of differentially expressed genes and regulatory transcription factors associated with chemoresistance.

METHODS: The study used a C57Bl/6 mouse model of B16 melanoma. The animals were randomly allocated into a control group and an experimental group, each with 12 mice. The experimental group was treated with dacarbazine at 50 mg/kg. Total RNA was extracted from tumor tissue, before high-throughput sequencing was performed. Data were subjected to bioinformatic processing for gene clustering and prediction of transcription factors for analyzing motifs in RNA sequences.

RESULTS: NGS sequencing and bioinformatic analysis yielded 670 differentially expressed genes, which were organized into 12 functional clusters related to DNA repair, apoptosis, and cell cycle. A comprehensive analysis identified key transcription factors (RELB, IRF5/7/4, NANOG, SOX2, LEF1, and NFKB2) that regulate signaling pathways essential for maintaining pluripotency (p = 0.000001), Wnt (p = 0.000753), TGF-β (p = 0.002631), Toll-like receptors (p = 0.000776), and NF-κB (p = 0.044609) associated with B16 melanoma cell resistance to the alkylating agent dacarbazine in vivo.

CONCLUSION: The findings of this study indicated that the epigenetic and transcription mechanisms of chemoresistance in melanoma cells involves maintenance of the stem cell phenotype, regulation of the immune response, and activation of the epithelial-mesenchymal transition.

Обоснование. Фенотипическая пластичность и гетерогенность клеток меланомы, обусловленные эпигенетической регуляцией и активностью транскрипционных факторов, играют ключевую роль в развитии резистентности к химиотерапии, что требует углублённого изучения молекулярных механизмов устойчивости к алкилирующим агентам, таким как дакарбазин.

Цель исследования. Анализ транскриптомных данных клеток меланомы B16 после воздействия дакарбазином in vivo для выявления ключевых кластеров дифференциально экспрессирующихся генов и регулирующих транскрипционных факторов, ассоциированных с резистентностью к терапии.

Методы. В исследовании использовали модель меланомы B16 у мышей C57Bl/6, разделённых на контрольную и опытную (дакарбазин в дозе 50 мг/кг) группы по 12 особей. Тотальную РНК экстрагировали из опухолевой ткани, проводили высокопроизводительное секвенирование с последующей биоинформатической обработкой для кластеризации генов и предсказания транскрипционных факторов — с использованием биоинформатических инструментов для анализа мотивов в последовательностях РНК.

Результаты. С помощью NGS-секвенирования и биоинформатического анализа выявлено 670 дифференциально экспрессирующихся генов, объединённых в 12 функциональных кластеров, связанных с репарацией ДНК, апоптозом и клеточным циклом. Идентифицированы ключевые транскрипционные факторы (RELB, IRF5/7/4, NANOG, SOX2, LEF1, NFKB2), регулирующие сигнальные пути поддержания плюрипотентности (p=0,000001), Wnt (p=0,000753), TGF-β (p=0,002631), Toll-подобных рецепторов (p=0,000776) и NF-κB (p=0,044609), ассоциированные с развитием устойчивости клеток меланомы B16 к алкилирующему агенту дакарбазину in vivo.

Заключение. Выявлены эпигенетические и транскрипционные механизмы устойчивости клеток меланомы к воздействию химиотерапевтических средств, включая поддержание стволового фенотипа, регуляцию иммунного ответа и активацию эпителиально-мезенхимального перехода.

论证：黑色素瘤细胞的表型可塑性和异质性，由表观遗传调控和转录因子活性决定，在化疗耐药性的发展中起着关键作用，这需要深入研究对达卡巴嗪等烷化剂产生耐药性的分子机制。

目的：分析体内暴露于达卡巴嗪后B16黑素瘤细胞的转录组学数据，以鉴定与治疗抗性相关的差异表达基因和调节转录因子的关键簇。方法：该研究在C57Bl/6小鼠中使用b16黑素瘤模型，分为对照组和实验组（剂量为50mg/kg的达卡巴嗪）12个个体。从肿瘤组织中提取总RNA，进行高通量测序，然后使用生物信息学工具分析RNA序列中的基序进行基因聚类和转录因子预测的生物信息学处理。

结果：NGS测序和生物信息学分析揭示了670个差异表达的基因分为12个与DNA修复、凋亡和细胞周期有关的功能簇。确定了调节维持多能性的信号传导途径的关键转录因子（RELB，IRF5/7/4、NANOG、SOX2、LEF1、NFKB2）、Wnt（p=0.000753）、TGF-β（p=0.002631）、Toll样受体（p=0.000776）和NF-kB（p=0.044609），与体内B16黑素瘤细胞对烷化剂达卡巴嗪的抗性发展有关。

结论：黑素瘤细胞对化疗剂影响的抗性的表观遗传和转录机制，包括维持茎表型、免疫应答的调节和上皮间充质过渡的激活。

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