METTL3对鼻咽癌细胞侵袭、转移及放射敏感性的影响  

Effect of METTL3 on invasion,metastasis and radiosensitivity of nasopharyngeal carcinoma cells

作　　者：刘滢滢 陈凯华[2] 孙永楚 覃月兰 宋阳光 朱小东 Liu Yingying;Chen Kaihua;Sun Yongchu;Qin Yuelan;Song Yangguang;Zhu Xiaodong(Department of Pathology,Wuming Hospital of Guangxi Medical University,Nanning 530100,China;Department of Radiation Oncology,Guangxi Medical University Cancer Hospital,Nanning 530021,China;Department of Oncology,Wuming Hospital of Guangxi Medical University,Nanning 530100,China;Department of Rehabilitation,Puyang Oilfield General Hospital,Puyang 457001,China;Guangxi Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor,Nanning 530021,China;Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor(Guangxi Medical University),Ministry of Education,Nanning 530021,China)

机构地区：[1]广西医科大学附属武鸣医院病理科,南宁530100 [2]广西医科大学附属肿瘤医院放疗科,南宁530021 [3]广西医科大学武鸣医院肿瘤科,南宁530100 [4]濮阳市油田总医院康复科,濮阳457001 [5]广西区域性高发肿瘤早期防治研究重点实验室,南宁530021 [6]区域性高发肿瘤早期防治研究教育部重点实验室(广西医科大学),南宁530021

出　　处：《中华放射肿瘤学杂志》2025年第2期167-175,共9页Chinese Journal of Radiation Oncology

基　　金：广西重点研发计划(桂科AB23026020);广西区域性高发肿瘤早期防治研究重点实验室项目(GKE-ZZ202230);南宁市武鸣区科学研究与技术开发计划项目(20230116)。

摘　　要：目的探讨甲基转移酶样蛋白3(METTL3)在鼻咽癌细胞CNE2、CNE-2R中的表达及其对细胞侵袭、转移能力及放射敏感性的机制研究。方法采用实时反转录PCR(RT-qPCR)和蛋白质印记法检测METTL3在正常鼻咽上皮细胞NP69和鼻咽癌细胞CNE2、鼻咽癌放射抵抗细胞CNE-2R中的表达水平。通过慢病毒介导的RNA干扰技术沉默鼻咽癌细胞中METTL3。采用划痕实验及Transwell实验检测细胞的迁移和侵袭能力。通过细胞克隆形成实验和CCK-8实验检测细胞的增殖能力及在不同剂量(0 Gy、2 Gy、4 Gy、6 Gy、8 Gy)X线照射后的细胞活力。用流式细胞术检测细胞凋亡情况。采用RNA甲基化免疫共沉淀(Me-RIP)实验检测沉默METTL3后,鼻咽癌细胞c-Jun的m6A修饰水平差异。通过放线菌素D实验检测沉默METTL3后细胞中c-Jun的转录稳定性。通过构建裸鼠移植瘤模型检测METTL3在体内对鼻咽癌放射敏感性的影响。结果与NP69细胞相比,METTL3在CNE2细胞中的mRNA和蛋白表达水平显著升高,在CNE-2R细胞中表达更高(均P<0.01)。使用慢病毒介导的RNA干扰技术成功构建稳定沉默METTL3的鼻咽癌细胞株(均P<0.01)。划痕实验及Transwell实验结果表明,沉默METTL3后CNE2、CNE-2R细胞的迁移、侵袭能力显著下降(均P<0.05)。克隆形成实验结果表明,经不同剂量X线照射后,沉默METTL3显著降低了CNE2、CNE-2R细胞的克隆形成集落数及存活分数(均P<0.05)。CCK-8实验结果表明,沉默METTL3的CNE2、CNE-2R细胞增殖能力明显降低(均P<0.05);经不同剂量照射后,沉默METTL3显著降低CNE2、CNE-2R细胞的存活分数(均P<0.05)。流式细胞术实验结果显示,在0 Gy和8 Gy照射剂量下,沉默METTL3后细胞凋亡率均高于对照组(均P<0.05)。Me-RIP实验结果表明,沉默METTL3后鼻咽癌细胞中c-Jun的m6A修饰水平显著降低(均P<0.01)。放线菌素D实验表明,沉默METTL3的鼻咽癌细胞中c-Jun转录稳定性下降。裸鼠移植瘤实验结果表明,�Objective To investigate the expression level of methyltransferase-like 3(METTL3)in nasopharyngeal carcinoma cells CNE2 and CNE-2R,and to evaluate the effect of METTL3 on cell invasion,metastasis and radiosensitivity.Methods Real-time reverse transcription PCR and Western blot were used to detect the expression levels of METTL3 in normal nasopharyngeal epithelial cells NP69 and nasopharyngeal carcinoma cells CNE2 and nasopharyngeal carcinoma radioresistant cells CNE-2R cells.METTL3 in CNE2 and CNE-2R cells was silenced by lentivirus-mediated RNA interference technology.The metastasis and invasion abilities of the cells were detected by the scratch assay and Transwell assay.Clonogenic assay and CCK-8 assay were employed to detect the proliferation capacity and viability of cells irradiated with different X-ray doses(0,2,4,6 and 8 Gy).Apoptosis was detected by flow cytometry.Methylated RNA immunoprecipitation(Me-RIP)assay was used to detect the difference in m6A modification level of c-Jun in CNE2 and CNE-2R cells after METTL3 silencing.The transcriptional stability of c-Jun in cells after silencing METTL3 was detected by actinomycin D assay.A nude mouse xenograft model was constructed to detect the effect of METTL3 on the radiosensitivity of nasopharyngeal carcinoma in vivo.Results Compared with NP69 cells,the expression levels of METTL3 mRNA and protein were significantly increased in CNE2 cells,and the expression level was even higher in CNE-2R cells(all P<0.01).Lentivirus-mediated RNA interference technology was used to construct a stable METTL3-silencing CNE2 and CNE-2R cell lines(both P<0.01).Scratch assay and Transwell assay showed that the metastasis and invasion abilities of CNE2 and CNE-2R cells were decreased significantly after METTL3 silencing(all P<0.05).Clonogenic assay showed that silencing METTL3 significantly reduced the number of colonies and survival fraction of CNE2 and CNE-2R cells after irradiation with different doses of X-rays(all P<0.05).CCK-8 assay showed that the proliferation ability of

关 键 词：鼻咽癌 METTL3 C-JUN 侵袭 肿瘤转移 放射敏感性 

分 类 号：R739.63[医药卫生—肿瘤]