Cui Xiao, Cao Chengcheng, Li Xinyang, Lin Biyan, Yan Aihui, Yang Ying
Department of Otorhinolaryngology, The First Hospital of China Medical University, Shenyang 110003, China.
Department of Pathology, Shengjing Hospital of China Medical University, Shenyang 110004, China.
Transl Oncol. 2024 Aug;46:102006. doi: 10.1016/j.tranon.2024.102006. Epub 2024 May 31.
The aggressive and refractory extranodal natural killer/T-cell lymphoma, nasal type (ENKTL-NT) is a subtype of non-Hodgkin's lymphoma. Succinylation promotes progression in a variety of tumors, but its mechanism in ENKTL-NT is unclear.
Bioinformatic analysis was performed to screen differentially expressed genes in the ENKTL dataset. Cell transfection techniques were used for knockdown and overexpression of genes. The mRNA and protein expression were detected using RT-qPCR and western blot, respectively. Immunohistochemical staining was used to assess protein expression in situ. For the detection of cell proliferation activity, CCK-8, clonal formation, and EDU staining assays were used. Flow cytometry was employed to detect apoptosis. Co-immunoprecipitation was utilized for the identification of protein interactions and succinylation modifications.
Succinyltransferase CPT1A was highly elevated in ENKTL-NT and was associated with a dismal prognosis. CPT1A knockdown suppressed SNK-6 cells' proliferation and induced apoptosis, while these effects were reversed by the overexpression of 14-3-3theta. Co-immunoprecipitation results showed that CPT1A caused succinylation of 14-3-3theta at site of K85, thereby enhancing the protein stability. Suppression of CPT1A-induced succinylation of 14-3-3theta by ST1326 resulted in the inhibition of SNK-6 cell proliferation and increased apoptosis. Paclitaxel combined with knockdown of CPT1A significantly inhibited the proliferation of ENKTL-NT compared to paclitaxel alone.
CPT1A induces succinylation of 14-3-3theta at the K85 site, promoting ENKTL-NT proliferation. The anti-ENKTL activity of paclitaxel was improved when combined with CPT1A knockdown.
侵袭性难治性结外自然杀伤/T细胞淋巴瘤，鼻型（ENKTL-NT）是非霍奇金淋巴瘤的一种亚型。琥珀酰化促进多种肿瘤进展，但其在ENKTL-NT中的机制尚不清楚。
进行生物信息学分析以筛选ENKTL数据集中差异表达的基因。使用细胞转染技术对基因进行敲低和过表达。分别使用RT-qPCR和蛋白质印迹法检测mRNA和蛋白质表达。免疫组织化学染色用于原位评估蛋白质表达。为检测细胞增殖活性，使用CCK-8、克隆形成和EDU染色试验。采用流式细胞术检测细胞凋亡。利用免疫共沉淀鉴定蛋白质相互作用和琥珀酰化修饰。
琥珀酰转移酶CPT1A在ENKTL-NT中高度升高，且与不良预后相关。CPT1A敲低抑制SNK-6细胞增殖并诱导凋亡，而14-3-3θ过表达可逆转这些作用。免疫共沉淀结果显示，CPT1A导致14-3-3θ在K85位点发生琥珀酰化，从而增强蛋白质稳定性。ST1326抑制CPT1A诱导的14-3-3θ琥珀酰化导致SNK-6细胞增殖受抑制且凋亡增加。与单独使用紫杉醇相比，紫杉醇联合CPT1A敲低显著抑制ENKTL-NT的增殖。
CPT1A诱导14-3-3θ在K85位点发生琥珀酰化，促进ENKTL-NT增殖。紫杉醇与CPT1A敲低联合使用时，其抗ENKTL活性增强。
