Li Yingxiao, Huang Bu-Miin, Tao Chih-Chieh, Lan Yu-Yan
School of Medicine, College of Medicine, I-Shou University, Kaohsiung, Taiwan, R.O.C.
Department of Biomedical Sciences, National Chung Cheng University, Chia-Yi, Taiwan, R.O.C.
Anticancer Res. 2025 Jul;45(7):3117-3126. doi: 10.21873/anticanres.17675.
BACKGROUND/AIM: Although current treatment strategies for nasopharyngeal carcinoma (NPC) have improved, the prognosis for a subset of patients remains poor, largely due to distant metastasis. Resistance to cisplatin is one of the key factors contributing to this unfavorable clinical outcome. This study investigated whether human umbilical cord mesenchymal stem cells-derived exosomes (hUC MSC-Exos) exhibit intrinsic anticancer activity and whether they can enhance the cytotoxic effects of cisplatin in NPC cells.
The human NPC cell line NPC-TW01 was utilized. Cell viability, protein expression, cytokeratin 18 fragment concentration, mitochondrial membrane potential (MMP) alterations, and reactive oxygen species (ROS) generation were assessed using MTT assay, immunoblotting, ELISA, MMP assay, and ROS detection assay, respectively.
MTT assay results indicated that treatment with 1×10 particles/ml hUC MSC-Exos slightly reduced the viability of NPC-TW01 cells. Notably, cotreatment with hUC MSC-Exos and 12.5 μg/ml cisplatin significantly enhanced cisplatin-induced cytotoxicity. This combination markedly increased the release of cytokeratin 18 fragments and the expression of cleaved caspase-7, both established markers of apoptosis. Moreover, the combination treatment up-regulated the expression of Fas, cleaved caspase-8, cleaved caspase-9, cleaved caspase-3, and tBid, and induced MMP disruption. In addition, the cotreatment significantly elevated ROS levels. Treatment with the ROS scavenger N-acetylcysteine not only suppressed ROS production but also attenuated the levels of cytokeratin 18 fragments, Fas, cleaved caspase-8, cleaved caspase-9, and cleaved caspase-3 induced by the combined treatment in NPC-TW01 cells.
hUC MSC-Exos enhance cisplatin-induced cytotoxicity in NPC-TW01 cells through ROS-dependent activation of the Fas-associated apoptotic signaling pathway. These results suggest that hUC MSC-Exos may serve as a promising adjuvant to augment the therapeutic efficacy of cisplatin in NPC.
背景/目的：尽管目前鼻咽癌（NPC）的治疗策略已有所改善，但部分患者的预后仍然较差，这主要是由于远处转移所致。对顺铂耐药是导致这一不良临床结局的关键因素之一。本研究调查了人脐带间充质干细胞来源的外泌体（hUC MSC-Exos）是否具有内在抗癌活性，以及它们是否能增强顺铂对NPC细胞的细胞毒性作用。
使用人NPC细胞系NPC-TW01。分别采用MTT法、免疫印迹法、酶联免疫吸附测定（ELISA）、线粒体膜电位（MMP）测定法和活性氧（ROS）检测法评估细胞活力、蛋白质表达、细胞角蛋白18片段浓度、MMP改变和ROS生成情况。
MTT法结果表明，用1×10颗粒/毫升的hUC MSC-Exos处理可轻微降低NPC-TW01细胞的活力。值得注意的是，hUC MSC-Exos与12.5微克/毫升顺铂联合处理显著增强了顺铂诱导的细胞毒性。这种联合处理显著增加了细胞角蛋白18片段的释放以及凋亡的既定标志物——裂解的半胱天冬酶-7的表达。此外，联合处理上调了Fas、裂解的半胱天冬酶-8、裂解的半胱天冬酶-9、裂解的半胱天冬酶-3和截短型Bid（tBid）的表达，并诱导MMP破坏。此外，联合处理显著提高了ROS水平。用ROS清除剂N-乙酰半胱氨酸处理不仅抑制了ROS的产生，还减弱了联合处理诱导的NPC-TW01细胞中细胞角蛋白18片段、Fas、裂解的半胱天冬酶-8、裂解的半胱天冬酶-9和裂解的半胱天冬酶-3的水平。
hUC MSC-Exos通过ROS依赖性激活Fas相关凋亡信号通路增强顺铂对NPC-TW01细胞的细胞毒性作用。这些结果表明，hUC MSC-Exos可能作为一种有前景的佐剂来增强顺铂对NPC的治疗效果。
