过表达通过 TLR4/MyD88/NF-κB 信号通路促进自噬来抑制弥漫性大 B 细胞淋巴瘤的进展。

Overexpression Inhibits Diffuse Large B-Cell Lymphoma Progression by Promoting Autophagy through TLR4/MyD88/NF-κB Signaling Pathway.

机构信息

Suzhou Medical College of Soochow University, 215006 Suzhou, Jiangsu, China.

Cancer Center, Department of Hematology, Zhejiang Provincial People's Hospital (Affiliated People's Hospital), Hangzhou Medical College, 310014 Hangzhou, Zhejiang, China.

出版信息

Discov Med. 2024 Aug;36(187):1627-1640. doi: 10.24976/Discov.Med.202436187.149.

DOI:10.24976/Discov.Med.202436187.149

PMID:39190378

Abstract

BACKGROUND

Tumor necrosis factor alpha induced protein 3 (TNFAIP3) is reportedly to have significant implications for autophagy regulation in various cancers. The current study aimed to decipher the role and mechanism of TNFAIP3 in diffuse large B-cell lymphoma (DLBCL) by modulating autophagy.

METHODS

Information pertaining to the differential expression and prognostic role of in DLBCL was gleaned from the Gene Expression Omnibus (GEO) database. The expression levels in human DLBCL cells were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. Cell counting kit-8 (CCK-8) and colony formation assays were employed to determine cell proliferation. Transwell assay and flow cytometry were applied to detect cell migration and apoptosis, respectively. Immunofluorescence and transmission electron microscope were used for the assessment of cell autophagy. The levels of apoptotic markers (caspase-3, cleaved-caspase-3, Bcl-2 Associated X (Bax), and B cell lymphoma-2 (Bcl-2)), autophagy indicators (the ratio of microtubule-associated proteins 1A/1B light chain 3 II and I (LC3II/LC3I), Sequestosome (p62)), and pathway proteins (toll-like receptor 4 (TLR4), myeloid differentiation primary response 88 (MyD88), Transcription Factor NF-Kappa-B P65 Subunit (p65), and phosphorylated-p65 (p-p65)) were assessed via Western blotting. Immunohistochemistry was employed to detect Ki67 expression in tumor tissues.

RESULTS

expression in DLBCL samples was downregulated, correlating with poor prognosis. expression was also downregulated in DLBCL cells. It was found that impeded cell proliferation and migration, and enhanced apoptosis of OCI-LY3 cells. Intervention with autophagy inhibitor 3-methyladenine (3-MA) markedly reversed apoptosis of OCI-LY3 cells induced by . Besides, induced autophagy via modulating the TLR4/MyD88/nuclear factor kappa B (NF-κB) signaling pathway. experiments showed that expression in DLBCL was downregulated, and upregulation of could inhibit tumor growth.

CONCLUSION

inhibits DLBCL progression by inducing TLR4/MyD88/NF-κB pathway-mediated autophagy.

摘要

背景

肿瘤坏死因子α诱导蛋白 3（TNFAIP3）据报道在各种癌症的自噬调控中具有重要意义。本研究旨在通过调节自噬来破译 TNFAIP3 在弥漫性大 B 细胞淋巴瘤（DLBCL）中的作用和机制。

方法

从基因表达综合数据库（GEO）中获取与 DLBCL 中差异表达和预后作用相关的信息。通过定量实时聚合酶链反应（qRT-PCR）和 Western blot 检测人 DLBCL 细胞中表达水平。细胞计数试剂盒-8（CCK-8）和集落形成实验用于检测细胞增殖。Transwell 实验和流式细胞术分别用于检测细胞迁移和凋亡。免疫荧光和透射电子显微镜用于评估细胞自噬。凋亡标志物（caspase-3、cleaved-caspase-3、Bcl-2 相关 X（Bax）和 B 细胞淋巴瘤-2（Bcl-2））、自噬标志物（微管相关蛋白 1A/1B 轻链 3 II 与 I（LC3II/LC3I）的比值、自噬相关蛋白（p62））和通路蛋白（Toll 样受体 4（TLR4）、髓样分化初级反应 88（MyD88）、转录因子 NF-κB P65 亚基（p65）和磷酸化-p65（p-p65））的水平通过 Western blot 进行评估。免疫组织化学用于检测肿瘤组织中 Ki67 的表达。

结果

在 DLBCL 样本中表达下调，与预后不良相关。在 DLBCL 细胞中也下调表达。结果发现，通过调节 TLR4/MyD88/核因子 kappa B（NF-κB）信号通路，抑制细胞增殖和迁移，增强 OCI-LY3 细胞凋亡。用自噬抑制剂 3-甲基腺嘌呤（3-MA）干预可显著逆转下调诱导的 OCI-LY3 细胞凋亡。此外，实验表明，DLBCL 中表达下调，上调可抑制肿瘤生长。