Russian Journal of PhysiologyRussian Journal of PhysiologyРоссийский физиологический журнал им. И.М. Сеченова0869-81392658-655XThe Russian Academy of Sciences67931410.31857/S0869813925020115UIEOFTEXPERIMENTAL ARTICLESЭКСПЕРИМЕНТАЛЬНЫЕ СТАТЬИResearch ArticleEffect of Chloroquine on Expression of Apoptosis and Autophagy Genes in MOLT-3 and IMR-32 CellsВлияние хлорохина на экспрессию генов апоптоза и аутофагии в клетках MOLT-3 и IMR-32ProkopenkoE. S.ПрокопенкоЕ. С.
Russian Federation
nagalak@mail.ruSokolovaT. V.СоколоваТ. В.
Russian Federation
nagalak@mail.ruNadeyO. V.НадейО. В.
Russian Federation
nagalak@mail.ruTrubnikovaA. D.ТрубниковаА. Д.
Russian Federation
nagalak@mail.ruAgalakovaN. I.АгалаковаН. И.
Russian Federation
nagalak@mail.ruSechenov Institute of Evolutionary Physiology and Biochemistry of the RASИнститут эволюционной физиологии и биохимии им. И.М. Сеченова РАНSaint Petersburg State UniversityСанкт-Петербургский государственный университет16022025111234936409052025Copyright ©; 2025, Russian Academy of SciencesCopyright ©; 2025, Российская академия наук2025Russian Academy of SciencesРоссийская академия наукhttps://journals.eco-vector.com/0869-8139/article/view/679314

The goal of the study was to compare an influence of chloroquine (CQ) on expression of apoptosis and autophagy genes in the cells of two tumor cells – leukemia MOLT-3 and neuroblastoma IMR-32 cultured in complete growth and serum-free RPMI-1640 and DMEM media, respectively, for 24 and 48 hours. The viability of cells was evaluated by MTT method, gene expression – by real time PCR. For MTT test, the cells were incubated with 10–100 µМ CQ. The expression of apoptosis (CASP3, BAX, BCL2) and autophagy (ULK1, BECN1, MAP1LC3B) genes was studied using 30 and 50 µМ CQ, which exerted considerable inhibitory effect on viability of cells of both lines, but did not promote their complete death. The sensitivity of both cell lines to CQ was higher in serum-free medium, however, the expression of apoptosis and autophagy genes substantially differed between them. In MOLT-3 cells, mRNA levels of pro-apoptotic genes CASP3 and BAX increased after 24-h incubation in serum-free medium, whereas in IMR-32 cells the expression of these genes increased only after 48-h in the presence of higher CQ concentration. In the cells of both lines 24-h CQ treatment resulted in enhanced expression of anti-apoptotic gene BCL2. In MOLT-3 cells, the absence of nutrients different combinations stimulated the genes of all three autophagy stages ULK1, BECN1 и MAP1LC3B, but none of applied treatment schemes did not affect the expression of ULK1 and MAP1LC3B genes in IMR-32 cells. Overall, 24-h culture with CQ under conditions of serum starvation appears to be more optimal for modulation of autophagy in MOLT-3 cells. In IMR-32 cells, CQ does not exert considerable influence on expression of autophagy genes, and their decreased viability is associated with activation of other mechanisms.

Целью исследования было сравнить влияние хлорохина (CQ) на экспрессию генов апоптоза и аутофагии в клетках двух опухолевых линий – лейкемии MOLT-3 и нейробластомы IMR-32, культивируемых в полных ростовых и бессывороточных средах RPMI-1640 и DMEM соответственно в течение 24 и 48 ч. Жизнеспособность клеток оценивали методом МТТ, экспрессию генов – методом ПЦР в реальном времени. Для теста МТТ клетки инкубировали с 10–100 мкМ CQ. Для изучения экспрессии генов апоптоза (CASP3, BAX, BCL2) и аутофагии (ULK1, BECN1, MAP1LC3B) использовали 30 и 50 мкМ CQ, которые оказывали значительный ингибирующий эффект на жизнеспособность клеток обеих линий, но не приводили к их полной гибели. Чувствительность клеток обеих линий к CQ была выше в бессывороточной среде, однако экспрессия генов апоптоза и аутофагии в них сильно различалась. В клетках MOLT-3 уровень мРНК проапоптотических генов CASP3 и BAX увеличивался после 24 ч инкубации в бессывороточной среде, в то время как в клетках IMR-32 повышение экспрессии этих генов наблюдалось только после 48 ч в присутствии более высокой концентрации CQ. В клетках обеих линий после 24-часовой обработки CQ увеличивалась экспрессия антиапоптотического гена BCL2. В клетках MOLT-3 в условиях недостатка питательных веществ наблюдались различные комбинации стимуляции генов всех трех этапов аутофагии ULK1, BECN1 и MAP1LC3B, но ни одна из схем обработки не оказала влияния на экспрессию генов ULK1 и MAP1LC3B в клетках IMR-32. Таким образом, 24-часовая обработка CQ в условиях сывороточного голодания является более оптимальной для модулирования аутофагии в клетках MOLT-3. В клетках IMR-32 CQ не оказывает значительного влияния на экспрессию генов аутофагии, а снижение их жизнеспособности связано с активацией других механизмов.

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