Department of Hematology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou City, Zhejiang Province 310016, PR China.
Department of Neurosurgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou City, Zhejiang Province 310016, PR China.
Cell Signal. 2024 Nov;123:111356. doi: 10.1016/j.cellsig.2024.111356. Epub 2024 Aug 22.
Diffuse large B-cell lymphoma (DLBCL) is the most common diagnosed subtype of lymphoma with high invasiveness and heterogeneity. Glycolysis is involved in regulating DLBCL progression. We aimed to explore the role of forkhead box protein A1 (FOXA1) in DLBCL and the mechanisms related to sirtuine5 (SIRT5) and glycolysis. FOXA1 expression in DLBCL cells was analyzed. Then, the proliferation and apoptosis of DLBCL cells were detected using Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EDU) staining and flow cytometry analysis following FOXA1 or SIRT5 knockdown. The glycolysis was assessed by measuring extracellular acidification rate (ECAR), glucose consumption and lactate secretion. Immunoblotting was employed to examine the expression of apoptosis- and glycolysis-related proteins. Additionally, luciferase reporter assay and chromatin immunoprecipitation (ChIP) assay were conducted to test the combination of FOXA1 to SIRT5 promotor region. Subsequently, SIRT5 expression was upregulated to conduct rescue assays. Finally, the effects of FOXA1 downregulation on the growth and glycolysis in OCI-ly7 tumor-bearing mice were examined. As a result, FOXA1 was upregulated in DLBCL cells and FOXA1 or SIRT5 knockdown inhibited the proliferation, accelerated the apoptosis and suppressed glycolysis reprograming in DLBCL cells. Importantly, FOXA1 could transcriptionally activate SIRT5 expression in DLBCL cells. Besides, SIRT5 overexpression counteracted the effects of FOXA1 deficiency on the proliferation, apoptosis and glycolysis reprogramming in DLBCL cells. Furthermore, FOXA1 knockdown inhibited the tumor growth, suppressed the glycolysis reprogramming and downregulated SIRT5 expression in vivo. In summary, FOXA1 could transcriptionally activate SIRT5 to reprogram glycolysis, thereby facilitating the malignant progression of DLBCL.
弥漫性大 B 细胞淋巴瘤（DLBCL）是最常见的淋巴瘤亚型，具有高侵袭性和异质性。糖酵解参与调节 DLBCL 的进展。我们旨在探讨叉头框蛋白 A1（FOXA1）在 DLBCL 中的作用以及与 SIRT5 和糖酵解相关的机制。分析了 DLBCL 细胞中的 FOXA1 表达。然后，通过 FOXA1 或 SIRT5 敲低后使用细胞计数试剂盒-8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EDU）染色和流式细胞术分析检测 DLBCL 细胞的增殖和凋亡。通过测量细胞外酸化率（ECAR）、葡萄糖消耗和乳酸分泌来评估糖酵解。免疫印迹用于检查凋亡和糖酵解相关蛋白的表达。此外，进行了荧光素酶报告基因测定和染色质免疫沉淀（ChIP）测定，以测试 FOXA1 与 SIRT5 启动子区域的结合。随后上调 SIRT5 表达进行挽救实验。最后，检查 FOXA1 下调对 OCI-ly7 荷瘤小鼠生长和糖酵解的影响。结果表明，FOXA1 在 DLBCL 细胞中上调，FOXA1 或 SIRT5 敲低抑制 DLBCL 细胞的增殖，加速凋亡并抑制糖酵解重编程。重要的是，FOXA1 可以在 DLBCL 细胞中转录激活 SIRT5 的表达。此外，SIRT5 过表达抵消了 FOXA1 缺乏对 DLBCL 细胞增殖、凋亡和糖酵解重编程的影响。此外，FOXA1 敲低抑制肿瘤生长，抑制体内糖酵解重编程并下调 SIRT5 表达。总之，FOXA1 可以转录激活 SIRT5 以重新编程糖酵解，从而促进 DLBCL 的恶性进展。
