Supplementary file 2_tRF-1:28-Val-CAC-2 promotes the development of nasopharyngeal cancer by targeting EPHB2.doc

IntroductionNasopharyngeal carcinoma (NPC) is highly aggressive, with a particularly high incidence in South China. is an aggressive cancer that affects particularly high numbers of patients in southern China and Southeast Asia. The cure rate of previous treatments is decreasing year by year, underscoring the need to devise new approaches to treating affected patients. This study was developed to examine the tRF-1:28-Val-CAC-2 expression in NPC and to elucidate its effects on proliferative, migratory, and apoptotic dynamics in NPC cells.
MethodsRT- qPCR was used to quantify tRF-1:28-Val-CAC-2 expression in NPC cells. Transfection was used to manipulate tRF-1:28-Val-CAC-2 expression levels, and proliferation, migration, and invasion were then evaluated through CCK-8, wound-healing, colony formation, and Transwell approaches. Apoptotic induction and cell cycle progression were assessed through flow cytometry, while EMT-related marker expression was assessed via qPCR and Western immunoblotting. The effects of tRF-1:28-Val-CAC-2 on the growth and distant metastasis of tumors were then tested in vivo using nude mice.
ResultsNPC cells exhibited tRF-1:28-Val-CAC-2 upregulation that was associated with significantly increased proliferative, migratory, and invasive activity together with the suppression of apoptotic death. In vivo experiments further confirmed the ability of tRF-1:28-Val-CAC-2 to promote tumor growth and distant metastasis. At a mechanistic level, these effects were related to the control of EPHB2 gene expression by tRF-1:28 Val-CAC-2, thereby shaping the survival and malignancy of the cells.
DiscussionThese results demonstrate that tRF-1:28-Val-CAC-2 promoted EPHB2 to enhance tumorigenic behavior in NPC cells, underscoring its key role as a novel target for therapeutic intervention.

引言：鼻咽癌（Nasopharyngeal carcinoma, NPC）是一种侵袭性极强的恶性肿瘤，在华南地区及东南亚发病率居高。现有治疗手段的治愈率逐年下降，凸显了开发新型治疗方案以改善患者预后的迫切性。本研究旨在检测鼻咽癌细胞中tRF-1:28-Val-CAC-2的表达水平，并阐明其对鼻咽癌细胞增殖、迁移及凋亡动态的调控作用。
方法：采用实时定量逆转录聚合酶链反应（RT-qPCR）定量检测鼻咽癌细胞中tRF-1:28-Val-CAC-2的表达水平；通过转染技术调控该分子的表达水平，随后分别采用CCK-8法、划痕愈合实验、平板克隆形成实验及Transwell实验评估细胞增殖、迁移与侵袭能力；采用流式细胞术检测细胞凋亡诱导情况及细胞周期进程，通过实时定量聚合酶链反应（qPCR）与蛋白质免疫印迹（Western immunoblotting）分析上皮间质转化（Epithelial-Mesenchymal Transition, EMT）相关标志物的表达水平；最后利用裸鼠体内模型验证tRF-1:28-Val-CAC-2对肿瘤生长及远处转移的影响。
结果：鼻咽癌细胞中tRF-1:28-Val-CAC-2呈现表达上调，且该上调与细胞增殖、迁移及侵袭能力显著增强、细胞凋亡受到抑制密切相关。体内实验进一步证实，tRF-1:28-Val-CAC-2可促进肿瘤生长与远处转移。机制分析显示，上述效应与tRF-1:28-Val-CAC-2对EPHB2基因表达的调控密切相关，进而影响鼻咽癌细胞的存活与恶性表型。
讨论：本研究结果表明，tRF-1:28-Val-CAC-2通过调控EPHB2基因表达增强鼻咽癌细胞的致瘤特性，凸显其作为新型治疗靶点的关键价值。