Abstract：
Diffuse large B-cell lymphoma (DLBCL) is characterized by significant treatment resistance. Palmitic acid (PA) has shown promising antitumor properties. This study aims to elucidate the molecular mechanisms by which PA influences DLBCL progression. We quantified the expression levels of microRNAs (miRNAs), Forkhead box protein O1 (FOXO1), and DNA methyltransferase 3A (DNMT3A) in both untreated and PA-treated DLBCL tumors and cell lines. Assessments were made of cell viability, apoptosis, and autophagy-related protein expression following PA administration. Interaction analyses among miR-429, DNMT3A, and FOXO1 were conducted using luciferase reporter assays and methylation-specific (MSP) Polymerase chain reaction (PCR). After transfecting the miR-429 inhibitor, negative control (NC) inhibitor, shRNA against DNMT3A (sh-DNMT3A), shRNA negative control (sh-NC), overexpression vector for DNMT3A (oe-DNMT3A), or overexpression negative control (oe-NC), we evaluated the effects of miR-429 and DNMT3A on cell viability, mortality, and autophagy-related protein expression in PA-treated DLBCL cell lines. The efficacy of PA was also tested in vivo using DLBCL tumor-bearing mouse models. MiR-429 and FOXO1 expression levels were downregulated, whereas DNMT3A was upregulated in DLBCL compared to the control group. PA treatment was associated with enhanced autophagy, mediated by the upregulation of miR-429 and downregulation of DNMT3A. The luciferase reporter assay and MSP confirmed that miR-429 directly inhibits DNMT3A, thereby reducing FOXO1 methylation. Subsequent experiments demonstrated that PA promotes autophagy and inhibits DLBCL progression by upregulating miR-429 and modulating the DNMT3A/FOXO1 axis. In vivo PA significantly reduced the growth of xenografted tumors through its regulatory impact on the miR-429/DNMT3A/FOXO1 axis. Palmitic acid may modulate autophagy and inhibit DLBCL progression by targeting the miR-429/DNMT3A/FOXO1 signaling pathway, suggesting a novel therapeutic target for DLBCL management.
摘要：
弥漫性大B细胞淋巴瘤(DLBCL)的特征在于显著的治疗抗性。棕榈酸（PA）已显示出有希望的抗肿瘤特性。本研究旨在阐明PA影响DLBCL进展的分子机制。我们量化了微小RNA（miRNA）的表达水平，叉头盒蛋白O1（FOXO1），和DNA甲基转移酶3A(DNMT3A)在未处理和PA处理的DLBCL肿瘤和细胞系中。对细胞活力进行评估，凋亡，和PA给药后自噬相关蛋白的表达。miR-429，DNMT3A，和FOXO1使用荧光素酶报告基因测定和甲基化特异性(MSP)聚合酶链反应(PCR)进行。转染miR-429抑制剂后，阴性对照（NC）抑制剂，针对DNMT3A的shRNA（sh-DNMT3A），shRNA阴性对照(sh-NC),DNMT3A的过表达载体（oe-DNMT3A），或过表达阴性对照（oe-NC），我们评估了miR-429和DNMT3A对细胞活力的影响，死亡率,和自噬相关蛋白在PA处理的DLBCL细胞系中的表达。还使用DLBCL荷瘤小鼠模型在体内测试了PA的功效。MiR-429和FOXO1表达水平下调，而DNMT3A在DLBCL中与对照组相比上调。PA治疗与自噬增强有关，由miR-429的上调和DNMT3A的下调介导。荧光素酶报告基因测定和MSP证实miR-429直接抑制DNMT3A,从而减少FOXO1甲基化。随后的实验表明，PA通过上调miR-429和调节DNMT3A/FOXO1轴来促进自噬并抑制DLBCL进展。体内PA通过其对miR-429/DNMT3A/FOXO1轴的调节作用显著降低异种移植肿瘤的生长。棕榈酸可能通过靶向miR-429/DNMT3A/FOXO1信号通路调节自噬并抑制DLBCL进展，提示DLBCL管理的新治疗目标。
