南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (12): 1783-1790.doi: 10.12122/j.issn.1673-4254.2022.12.05

• • 上一篇    下一篇

PRMT1通过促进RRM2表达抑制鼻咽癌细胞凋亡

周兰柱，吴 俊，张明洁 ，赵 报 ，马士崟   

  1. 蚌埠医学院第一附属医院耳鼻咽喉头颈外科，安徽 蚌埠 233000
  • 出版日期:2022-12-20 发布日期:2023-01-12

PRMT1 inhibits apoptosis of nasopharyngeal carcinoma cells by promoting RRM2 expression

ZHOU Lanzhu, WU Jun, ZHANG Mingjie, ZHAO Bao, MA Shiyin   

  1. Department of Otorhinolaryngology-Head and Neck Surgery, First Affiliated Hospital of Bengbu Medical College, Bengbu 233000, China
  • Online:2022-12-20 Published:2023-01-12

RichHTML

42

PDF

404

摘要： 目的 探讨PRMT1能否通过促进RRM2抑制鼻咽癌细胞凋亡。方法 免疫组化和Western blot共同检测鼻咽癌和癌旁组织中PRMT1及RRM2的相对表达量；以人正常鼻黏膜上皮细胞系（HNEpC）作为对照，Western blot实验检测不同鼻咽癌细胞系中PRMT1和RRM2蛋白的相对表达量；对CNE-2细胞中PRMT1分别进行过表达和下调，过表达实验设阴性对照组（oe NC：CNE-2细胞中转染pcDNA3.1质粒）、过表达PRMT1组（oe PRMT1：CNE-2细胞中转染pcDNA3.1 PRMT1质粒）、下调实验设阴性对照组（si-NC：CNE-2细胞中转染NC无关序列）及PRMT1小干涉RNA组（si-PRMT1：CNE-2细胞中转染PRMT1小干涉RNA），对CNE-2细胞中RRM2分别进行过表达和下调，过表达实验设阴性对照组（oe NC：CNE-2细胞中转染pcDNA3.1质粒）、过表达RRM2组（oe RRM2：CNE-2细胞中转染pcDNA3.1 RRM2质粒）、下调实验设阴性对照组（si-NC：CNE-2细胞中转染NC无关序列）及RRM2小干涉RNA组（si-RRM2：CNE-2细胞中转染RRM2小干涉RNA）。Western blot验证PRMT1和RRM2的表达关系；AnnexinV-FITC/PI凋亡检测试剂盒检测细胞凋亡；活性氧试剂盒检测细胞活性氧。结果 与癌旁组织和HNEpC细胞相比，鼻咽癌组织和CNE-2细胞中的PRMT1和RRM2相对表达量均显著增加（P<0.05）；Western blot证明PRMT1可促进RRM2表达，过表达PRMT1或RRM2，CNE-2细胞活性氧产生量与凋亡率均降低（P<0.05），下调PRMT1或RRM2表达，CNE-2细胞活性氧产生量与凋亡率凋亡率均增加（P<0.05）。Western blot显示，过表达PRMT1或RRM2表达，Cleaved caspase-3和Cleaved caspase-8蛋白表达减少（P<0.05），下调PRMT1或RRM2表达，Cleaved caspase-3和Cleaved caspase-8蛋白表达增加（P<0.05）；当同时下调PRMT1和过表达RRM2表达，与单独下调PRMT1组比，CNE-2细胞活性氧产生量与凋亡率均显著降低（P<0.05），Cleaved caspase-3和Cleaved caspase-8蛋白表达均减少（P均<0.05）。结论 PRMT1通过促进RRM2表达影响CNE-2细胞凋亡。

关键词: 鼻咽癌；PRMT1；RRM2；凋亡

Abstract: Objective To verify whether PRMT1 inhibits apoptosis of nasopharyngeal carcinoma (NPC) cells by promoting RRM2 expression. Methods Immunohistochemistry and Western blotting were performed to detect the relative expression of PRMT1 and RRM2 in NPC and adjacent tissues and in different NPC cell lines and a normal nasal mucosal epithelial cell line (HNEpC). Experiments of PRMT1 or RRM2 overexpression or siRNA-mediated PRMT1 or RRM2 knockdown were carried out in CNE-2 cells to investigate the relationship between PRMT1 and RRM2 expressions using Western blotting. Apoptosis of the transfected cells was detected using Annexin V-FITC/PI apoptosis detection kit, and the production of intracellular reactive oxygen species (ROS) was determined using a ROS detection kit. Results Compared with adjacent tissues and HNEpC cells, NPC tissues and cell lines expressed significantly higher levels of PRMT1 and RRM2 (P<0.05). In CNE-2 cells with PRMT1 or RRM2 overexpression or knockdown, Western blotting demonstrated that PRMT1 could positively regulate the expression of RRM2 (P<0.05). Overexpression of PRMT1 or RRM2 significantly reduced intracellular ROS production and apoptosis rate of CNE-2 cells (P<0.05), and PRMT1 or RRM2 knockdown strongly increased ROS production and cell apoptosis (P<0.05). Overexpression of either PRMT1 or RRM2 significantly decreased the expressions of cleaved caspase-3 and cleaved caspase-8 proteins (P<0.05), and PRMT1 or RRM2 knockdown obviously promoted their expressions (P<0.05). PRMT1 knockdown combined with RRM2 overexpression, as compared with PRMT1 knockdown only, significantly decreased ROS production and cell apoptosis (P<0.05) as well as the protein expressions of cleaved caspase-3 and cleaved caspase-8 in CNE-2 cells (P<0.05). Conclusion The high expression of PRMT1 in NPC inhibits apoptosis of NPC cells by promoting the expression of RRM2.

Key words: nasopharyngeal carcinoma; PRMT1; RRM2; apoptosis