Effect of Chloroquine on Expression of Apoptosis and Autophagy Genes in MOLT-3 and IMR-32 Cells
- Authors: Prokopenko E.S.1,2, Sokolova T.V.1, Nadey O.V.1, Trubnikova A.D.1, Agalakova N.I.1
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Affiliations:
- Sechenov Institute of Evolutionary Physiology and Biochemistry of the RAS
- Saint Petersburg State University
- Issue: Vol 111, No 2 (2025)
- Pages: 349-364
- Section: EXPERIMENTAL ARTICLES
- URL: https://edgccjournal.org/0869-8139/article/view/679314
- DOI: https://doi.org/10.31857/S0869813925020115
- EDN: https://elibrary.ru/UIEOFT
- ID: 679314
Cite item
Abstract
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.
Keywords
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About the authors
E. S. Prokopenko
Sechenov Institute of Evolutionary Physiology and Biochemistry of the RAS; Saint Petersburg State University
Author for correspondence.
Email: nagalak@mail.ru
Russian Federation, Saint Petersburg; Saint Petersburg
T. V. Sokolova
Sechenov Institute of Evolutionary Physiology and Biochemistry of the RAS
Email: nagalak@mail.ru
Russian Federation, Saint Petersburg
O. V. Nadey
Sechenov Institute of Evolutionary Physiology and Biochemistry of the RAS
Email: nagalak@mail.ru
Russian Federation, Saint Petersburg
A. D. Trubnikova
Sechenov Institute of Evolutionary Physiology and Biochemistry of the RAS
Email: nagalak@mail.ru
Russian Federation, Saint Petersburg
N. I. Agalakova
Sechenov Institute of Evolutionary Physiology and Biochemistry of the RAS
Email: nagalak@mail.ru
Russian Federation, Saint Petersburg
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