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Archive > Volume 33 Issue 3 > 2026,33(3):243-251. DOI:10.3872/j.issn.1007-385X.2026.03.003 Prev Next
Basic Research
Study on the role of ACTL6A in promoting doxorubicin resistance in diffuse large B cell lymphoma cells by regulating GPX4-mediated ferroptosis
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Abstract:
[Abstract] Objective: To investigate the mechanism by which actin-like protein 6A (ACTL6A) regulates ferroptosis and contributes to doxorubicin (DOX) resistance in diffuse large B cell lymphoma (DLBCL) cells. Methods: The parental DLBCL cell line SU-DHL-4 cells and its DOX-resistant variant SU-DHL-4/DOX cells were cultured. Changes in the expression of ACTL6A were detected by qPCR and WB assay. SU-DHL-4/DOX cells with ACTL6A knockdown were constructed by transfecting plasmids carrying a short hairpin RNA targeting ACTL6A (sh-ACTL6A) or its negative control (sh-NC). The expression levels of ACTL6A and ferroptosis-related proteins, including glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), and acyl-CoA synthetase longchain family member 4 (ACSL4), were measured using qPCR and WB. Chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays were performed to verify the targeting and regulatory relationship between ACTL6A and GPX4. SU-DHL-4/DOX cells were divided into Control, sh-NC, sh-ACTL6A, sh-NC + oe-GPX4, and sh-ACTL6A + oe-GPX4 groups. Corresponding plasmids were transfected into the cells. CCK-8 assay was used to detect cell survival rates of each group under different concentrations of DOX treatment. FerroOrange, Liperfluo, and DCFH-DA probes were used to detect ferrous ion (Fe2+) levels, lipid peroxidation, and reactive oxygen species (ROS) in each group of cells, respectively. A colorimetric method was used to measure the contents of glutathione (GSH) and malondialdehyde (MDA) in each group of cells. Results: Both ACTL6A mRNA and protein were highly expressed in SU-DHL-4/DOX cells (both P < 0.05), compared to SU-DHL-4 cells. ACTL6A and GPX4 have a targeting binding relationship. Knockdown of ACTL6A significantly decreased the mRNA and protein expression of ACTL6A and GPX4 in SU-DHL-4/DOX cells (both P < 0.05), indicating that ACTL6A regulates GPX4 expression. Knockdown of ACTL6A significantly inhibited the survival of SU-DHL-4/DOX cells, increased intracellular Fe2+, lipid peroxides, ROS, and MDA levels, and inhibited GSH production (all P < 0.05). However, overexpression of GPX4 in ACTL6A-knockdown cells upregulated the mRNA and protein expression levels of GPX4 in SU-DHL-4/DOX cells (both P < 0.05), increased cell survival rate, inhibited the production of intracellular Fe2+, lipid peroxides, ROS, and MDA, and increased GSH production (all P < 0.05). Conclusion: ACTL6A is highly expressed in DOX-resistant DLBCL cells. By regulating GPX4 expression, ACTL6A inhibits ferroptosis and promotes drug resistance in DLBCL cells.
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