与GRP78相关的miR-15a-5p在鼻咽癌放疗抵抗中的作用OA
The role of miR-15a-5p associated with GRP78 in radiotherapy resistance of nasopharyngeal carcinoma
目的 探究与葡萄糖调节蛋白 78(GRP78)相关的非编码微小 RNA-15a-5p(miR-15a-5p)在鼻咽癌放疗抵抗中的作用并初步探索二者之间的调控机制.方法 应用 shRNA 技术干扰鼻咽癌细胞 GRP78 表达,通过miRNA 芯片筛选与 GRP78 相关 miRNA,并结合 RT-PCR 实验验证敲低后 miR-15a-5p 表达水平;在鼻咽癌细胞系中瞬转 miR-15a-5p 相应的模拟物和抑制剂后验证 miR-15a-5p 在鼻咽癌放疗抵抗中的作用;通过生物信息学初步分析 GRP78 调控 miR-15a-5p 的可能分子机制.结果 建立的鼻咽癌放射抵抗细胞系 C666-IR 对放射的抵抗性强于母本细胞 C666-1,且 GRP78 蛋白的表达更高,分布更广;C666-IR 细胞中 miR-15a-5p 的表达高于 C666-1 细胞,且GRP78 高表达的鼻咽癌细胞经射线照射后 miR-15a-5p 仍维持较高水平,而敲低 GRP78 可以降低鼻咽癌细胞的miR-15a-5p 水平;瞬转 miR-15a-5p 模拟物增强 C666-1细胞的迁移能力和放射抵抗性,瞬转 miR-15a-5p 抑制剂降低C666-IR 细胞的迁移能力和放射抵抗性;生物信息学分析预测 miR-15a-5p 基因启动子 460 位点左右区域与转录因子结合可能性最大,且预测到 13 个可能的转录因子,其中 STAT4 和 FOXP3 可能性最大.结论 GRP78 介导 miR-15a-5p 表达上调促进鼻咽癌放疗抵抗,其调控机制可能为 GRP78 通过驱动转录因子 STAT4/FOXP3 提高 miR-15a-5p 表达,靶向 miR-15a-5p 可能为放疗抵抗的鼻咽癌提供新的治疗策略.
Objective To investigate the role of non-coding microRNA-15a-5p(miR-15a-5p)associated with glucose-regulated protein 78(GRP78)in radioresistance of nasopharyngeal carcinoma and to preliminarily explore the regulatory mechanism between them.Methods The expression of GRP78 in nasopharyngeal carcinoma cells was interfered by shRNA technology.The miRNA related to GRP78 was screened by miRNA chip,and the expression level of miR-15a-5p after knockdown was verified by reverse transcription-polymerase chain reaction.The role of miR-15a-5p in nasopharyngeal carcinoma radiotherapy resistance was verified by transient transfection of miR-15a-5p corresponding mimics and inhibitors in nasopharyngeal carcinoma cell lines.The possible molecular mechanism of GRP78 regulating miR-15a-5p was preliminarily analyzed by bioinformatics.Results The established radioresistant nasopharyngeal carcinoma cell line C666-IR showed stronger resistance to radiation than its parental cell C666-1,and had higher expression and wider distribution of GRP78 protein.The expression of miR-15a-5p in C666-IR cells was higher than that in C666-1 cells.After irradiation,the level of miR-15a-5p remained high in nasopharyngeal carcinoma cells with high expression of GRP78,while knockdown of GRP78 could reduce the level of miR-15a-5p in nasopharyngeal carcinoma cells.Transient transfection of miR-15a-5p mimics enhanced the migration ability and radioresistance of C666-1 cells,while transient transfection of miR-15a-5p inhibitors reduced the migration ability and radioresistance of C666-IR cells.Bioinformatics analysis predicted that the region around the 460 site of the miR-15a-5p gene promoter was most likely to bind to transcription factors,and 13 possible transcription factors were predicted,of which STAT4 and FOXP3 had the greatest possibility.Conclusions GRP78 mediates the up-regulation of miR-15a-5p expression to promote radiotherapy resistance of nasopharyngeal carcinoma.The regulatory mechanism may be that GRP78 increases the expression of miR-15a-5p by driving the transcription factor STAT4/FOXP3.Targeting miR-15a-5p may provide a new therapeutic strategy for radiotherapy-resistant nasopharyngeal carcinoma.
莫妍;冯雪萍;吕武武
中南大学湘雅医院 国家老年疾病临床研究中心 肿瘤科/医学科学研究中心,湖南 长沙 410008中南大学湘雅医院 国家老年疾病临床研究中心 肿瘤科/医学科学研究中心,湖南 长沙 410008浙江大学医学院附属第四医院 肿瘤科,浙江 义乌 321000
医药卫生
鼻咽癌葡萄糖调节蛋白78微小RNA-15a-5p放疗抵抗
Nasopharyngeal carcinomaGRP78miR-15a-5pRadiotherapy resistance
《中国耳鼻咽喉颅底外科杂志》 2026 (2)
26-32,7
国家自然科学基金面上项目(82273486).
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