南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (10): 1665-1673.doi: 10.12122/j.issn.1673-4254.2023.10.03

• • 上一篇    下一篇

淫羊藿素通过调控铁死亡增加鼻咽癌细胞的放射敏感性

胡 桐，勾文峰，任中昊，刘改廷，李祎亮，左代英，侯文彬   

  1. 中国医学科学院放射医学研究所天津市放射医学与分子核医学重点实验室，天津 300192；沈阳药科大学生命科学与生物制药学院，辽宁 沈阳 110016
  • 出版日期:2023-10-20 发布日期:2023-11-02

Icaritin increases radiosensitivity of nasopharyngeal carcinoma cells by regulating iron death

HU Tong, GOU Wenfeng, REN Zhonghao, LIU Gaiting, LI Yiliang, ZUO Daiying, HOU Wenbin   

  1. Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Institute of Radiation Medicine, Chinese Academy of Medical Sciences, Tianjin 300192, China; School of Life Sciences and Biopharmaceutical, Shenyang Pharmaceutical University, Shenyang 110016, China
  • Online:2023-10-20 Published:2023-11-02

RichHTML

33

PDF

330

摘要： 目的 探索淫羊藿素对鼻咽癌细胞放射增敏作用以及机制。方法 通过MTT实验和克隆形成实验，观察淫羊藿素对鼻咽癌HONE1和HNE1细胞增殖能力的影响，筛选出给药剂量以及照射剂量。建立鼻咽癌细胞γ射线照射模型以及淫羊藿素给药模型，将细胞分为空白组、淫羊藿素组、照射组和淫羊藿素与照射联合组。通过流式细胞术检测4组细胞活性氧（ROS）的变化、周期分布以及细胞凋亡情况，通过Western blot实验检测DNA损伤标志蛋白γ-H2AX、周期相关蛋白CDC25C、p-CDC25C、和CyclinB1以及铁死亡标志蛋白ACSL4和GXP4的表达情况。构建HONE1细胞裸鼠皮下移植瘤模型，观察淫羊藿素与放射联合对肿瘤组织的生长情况的影响。结果 淫羊藿素以剂量依赖性方式抑制鼻咽癌细胞活力，增强放射对鼻咽癌细胞增殖能力的抑制作用。流式细胞术及Western blot实验显示，淫羊藿素与放射联合后促进鼻咽癌细胞ROS增多，γ-H2AX表达上调（P<0.001）。与单纯照射组相比，联合组会使鼻咽癌细胞发生G2期阻滞，CDC25C和CyclinB1表达下调，p-CDC25C表达上调（P<0.01）。联合组会促进鼻咽癌细胞发生细胞凋亡，铁死亡蛋白ACSL4表达上调而GXP4表达下调（P<0.001）。与空白组相比，其余3组的肿瘤生长速度明显减慢。与照射组相比，联合组肿瘤生长速度减慢，瘤体重量下降（P<0.001）。结论 淫羊藿素可在体内、体外均增强鼻咽癌对放射的敏感性，可能主要是通过增强细胞活性氧促进鼻咽癌细胞发生铁死亡来达到放射增敏的作用。

关键词: 淫羊藿素；鼻咽癌；放射增敏；活性氧；铁死亡

Abstract: Objective To explore the radiosensitizing effect of icaritin on nasopharyngeal carcinoma (NPC) cells and the underlying mechanism. Methods MTT assay and clonal formation assay were used to evaluate the effect of icaritin on proliferation of human NPC HONE1 and HNE1 cells. The effects of icaritin treatment, γ-ray radiation, or both on production of reactive oxygen species (ROS), cell cycle distribution and apoptosis of the NPC cells were assessed using flow cytometry. The expressions of DNA damage markers γ-H2AX, cycle-related proteins CDC25C, p-CDC25C and cyclin B1, and ferroptosis markers ACSL4 and GXP4 were detected using Western blotting. A nude mouse model bearing subcutaneous HONE1 cell xenograft was used to observe the effect of icaritin and radiation on tumor growth. Results Icaritin dose-dependently inhibited the viability of the NPC cells and enhanced the inhibitory effect of radiation on cell proliferation. Flow cytometry and Western blotting showed that icaritin treatment prior to radiation significantly promoted ROS production and γ-H2AX expression in the NPC cells (P<0.001). Compared with radiation exposure alone, the combined treatment caused cell cycle arrest in G2 phase, down-regulated CDC25C and cyclin B1 expression, and up-regulated p-CDC25C expression in the cells (P<0.01), resulting also in increased cell apoptosis, enhanced expression of ferroptosis protein ACSL4 and lowered expression of GXP4 (P<0.001). In the tumor-bearing mice, icaritin treatment, compared with radiation alone, significantly reduced the tumor growth rate and decreased tumor weight (P<0.001). Conclusion Icaritin can enhance radiosensitivity of NPC cells both in vitro and in nude mice possibly by enhancing ROS production to promote iron death of the cells.

Key words: icaritin; nasopharyngeal carcinoma; radiosensitization; reactive oxygen species; ferroptosis