miR-24对鼻咽癌放射敏感性的影响
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摘要
目的:探讨miR-24在鼻咽癌中的表达和对鼻咽癌细胞放射敏感性的影响。方法:收集广西医科大学第一附属医院82例鼻咽癌组织及22例正常鼻咽组织,基因芯片筛选鼻咽癌细胞中差异表达的miRNA谱;实时荧光定量PCR(qPCR)技术检测miR-24的表达,分析miR-24的表达水平与临床分期的关系及对疗效的影响。构建鼻咽癌细胞CNE1稳转株及鼻咽癌细胞CNE1miR-24裸鼠模型,MTT实验和克隆形成实验检测实验组(miR-24组)与空白对照组(miR-NC组)鼻咽癌细胞增殖率及不同照射剂量处理后的生存率;单剂量12Gy/f/d局部照射鼻咽癌裸鼠移植瘤后,比较两组裸鼠的肿瘤体积。结果:基因芯片结果显示miR-24表达下调最为明显;miR-24在鼻咽癌细胞CNE1低于正常鼻咽上皮细胞NP69(P<0.01);miR-24在T分期、N分期及临床分期的早期表达较高(P<0.01);低表达miR-24的鼻咽癌患者治疗无效率明显高于高表达患者(P<0.05);体外实验结果显示,与miR-NC组比较,miR-24组细胞增殖率及生存率明显降低(P<0.01或P<0.05);体内实验表明,与miR-NC组比较,照射后miR-24组移植瘤体积明显减小(P<0.001)。结论:过表达miR-24能够抑制鼻咽癌细胞的增殖,增加鼻咽癌细胞的放射敏感性,且高表达miR-24的鼻咽癌患者治疗效果较好。
Objective:To investigate the expression of miR-24 in nasopharyngeal carcinoma(NPC)patients and its effect on radiosensitivity in vitro and in vivo.Methods:The nasopharyngeal tissues from 82 NPC patients and 22 cases of healthy controls were collected and miR-24 expression in these tissues was detected by qPCR.The correlation between the expression of miR-24 and the clinical stage and efficacy was analyzed.The differentially expressed miRNA profiles in NPC cells were identified by microarray.The NPC CNE1 cells or BALB/C nude mice were divided into a highly-expressed miR-24 group and a control group(miRNC group).MTT assay and clone formation assay were used to detect cell proliferation rates and cloning efficiency.BALB/C nude mice subjected to NPC were treated with 12 Gy partial irradiation,and the tumor volume was compared between the two groups.Results:Microarray assay results showed that miR-24 expression was significantly downregulated in NPC cells.The expression of miR-24 in NPC tissues was higher than that in controls,and it was correlated with tumor size and clinical stage(P <0.01).The therapeutic effect in patients with high expression of miR-24 was superior to those with low-expression of miR-24(P <0.05).Compared with miR-NC group,the cell proliferation rate,survival rate and the transplanted tumor size were reduced in miR-24 group.Conclusion:Over expressed miR-24 could inhibit the proliferation of NPC cells and enhance radiosensitivity of NPCin vitro and in vivo.The NPC patients with highly-expressed miR-24 had better prognosis.
Objective:To investigate the expression of miR-24 in nasopharyngeal carcinoma(NPC)patients and its effect on radiosensitivity in vitro and in vivo.Methods:The nasopharyngeal tissues from 82 NPC patients and 22 cases of healthy controls were collected and miR-24 expression in these tissues was detected by qPCR.The correlation between the expression of miR-24 and the clinical stage and efficacy was analyzed.The differentially expressed miRNA profiles in NPC cells were identified by microarray.The NPC CNE1 cells or BALB/C nude mice were divided into a highly-expressed miR-24 group and a control group(miRNC group).MTT assay and clone formation assay were used to detect cell proliferation rates and cloning efficiency.BALB/C nude mice subjected to NPC were treated with 12 Gy partial irradiation,and the tumor volume was compared between the two groups.Results:Microarray assay results showed that miR-24 expression was significantly downregulated in NPC cells.The expression of miR-24 in NPC tissues was higher than that in controls,and it was correlated with tumor size and clinical stage(P <0.01).The therapeutic effect in patients with high expression of miR-24 was superior to those with low-expression of miR-24(P <0.05).Compared with miR-NC group,the cell proliferation rate,survival rate and the transplanted tumor size were reduced in miR-24 group.Conclusion:Over expressed miR-24 could inhibit the proliferation of NPC cells and enhance radiosensitivity of NPCin vitro and in vivo.The NPC patients with highly-expressed miR-24 had better prognosis.
引文
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[2]XU L,CHEN Z,XUE F,et al.MicroRNA-24inhibits growth,induces apoptosis,and reverses radioresistance in laryngeal squamous cell carcinoma by targeting Xlinked inhibitor of apoptosis protein[J].Cancer Cell International,2015,15(1):61.
[3]ZHAO G,LIU L,ZHAO T,et al.Upregulation of miR-24promotes cell proliferation by targeting NAIF1in non-small cell lung cancer[J].Tumour Biology,2015,36(5):3693-3701.
[4]MA Y,SHE XG,MING YZ,et al.miR-24promotes the proliferation and invasion of HCC cells by targeting SOX7[J].Tumour Biology,2014,35(11):10731-10736.
[5]DU WW,FANG L,LI M,et al.MicroRNA miR-24enhances tumor invasion and metastasis by targeting PTPN9and PTPRF to promote EGF signaling[J].Journal of Cell Science,2013,126(Pt 6):1440-1453.
[6]龙满美,黄小英,姚金光,等.microRNA-24表达与黄曲霉毒素B1相关性肝细胞癌预后的关联性研究[J].上海交通大学学报(医学版),2014,34(12):1743-1748.
[7]XU CX,XU M,TAN L,et al.MicroRNA miR-214regulates ovarian cancer cell stemness by targeting p53/Nanog[J].The Journal of Biological Chemistry,2012,287(42):34970-34978.
[8]PANDA H,CHUANG TD,LUO X,et al.Endometrial miR-181aand miR-98expression is altered during transition from normal into cancerous state and target PGR,PGRMC1,CYP19A1,DDX3X,and TIMP3[J].The Journal of Clinical Endocrinology and Metabolism,2012,97(7):E1316-E1326.