甲基转移酶METTL3对口腔鳞癌细胞放疗敏感性的调控作用研究  

Regulation of methyltransferase METTL3 on radiotherapy sensitivity of oral squamous cell carcinoma cells

作　　者：孟庆哲 黄军红 杨新杰 李欢 杨子桧 王珺 李雅慧 刘荣 魏建华 MENG Qingzhe;HUANG Junhong;YANG Xinjie;LI Huan;YANG Zihui;WANG Jun;LI Yahui;LIU Rong;WEI Jianhua(College of Stomatology,Jiamusi University,Affiliated Stomatological Hospital Heilongjiang Provincial Key Laboratory of Oral Biomedical Materials and Clinical Applications,154000,China;State Key Laboratory of Oral&Maxillofacial Reconstruction and Regeneration,National Clinical Research Center for Oral Diseases,Shaanxi Clinical Research Center for Oral Diseases,Department of Oral and Maxillofacial Surgery,School of Stomatology,The Fourth Military Medical University,Xi'an)

机构地区：[1]佳木斯大学附属口腔医学院·附属口腔医院黑龙江省口腔生物医学材料及临床应用重点实验室,154000 [2]口颌系统重建与再生全国重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔疾病临床医学研究中心,空军军医大学口腔医院口腔颌面外科

出　　处：《实用口腔医学杂志》2025年第2期206-213,共8页Journal of Practical Stomatology

基　　金：国家自然科学基金(编号:81973114)。

摘　　要：目的:研究甲基转移酶METTL3对口腔鳞癌细胞放疗敏感性的影响。方法:对口腔鳞癌细胞CAL27、SCC9和SCC15分别给予2、4和8 Gy的X射线照射,流式细胞术检测细胞的凋亡水平,qRT-PCR及Western blot检测不同放射剂量下细胞中甲基化基因的RNA及蛋白表达,LC-MS/MS法对细胞中m6A进行定量,流式细胞术检测敲低及过表达METTL3后CAL27及SCC15中细胞凋亡水平,克隆形成实验观察在CAL27及SCC15中敲低和过表达靶基因后进行放射处理,观测细胞克隆形成率的变化。结果:3种细胞凋亡率均呈X射线剂量依赖性升高,各剂量下CAL27细胞凋亡率最高,SCC15细胞最低。CAL27中METTL3的RNA及蛋白表达水平显著低于SCC15。m6A定量结果显示CAL27中甲基化修饰水平低于SCC15。CAL27及SCC15细胞经不同剂量放射处理后,METTL3的表达均升高。敲低METTL3后均使细胞的凋亡率增加及克隆形成率降低,过表达METTL3使细胞凋亡率减少、克隆形成率升高。结论:调控METTL3可以影响口腔鳞癌细胞的放射治疗敏感性,有可能成为口腔鳞癌放疗增敏的新靶点。Objective:To study the influence of methyltransferases like 3(METTL3)on the radiosensitivity of oral squamous cell carcinoma cells(OSCCs).Methods:The apoptosis level of OSCCs CAL27,SCC9 and SCC15 treated with X-ray radiation doses of 2,4 and 8 Gy respectively was compared by flow cytometry,the expression of methylated gene RNA and protein in the cells were examined by qRT-PCR and Western blot.m6A in the cells was quantified by LC/LC-MS method.qRT-PCR and Western blot were used to investigate the expression of methylated gene RNA and protein in the cells.Flow cytometry was used to examine the cell apoptosis level of CAL27 and SCC15 cells treated with METTL3 overexpression and knockdown respectively.The clone formation of CAL27 and SCC15 cells after knockdown and overexpression of target genes followed by radiation treatment was observed by clonogenic assays.Results:The apoptosis rate of all the cell lines increased with the increase dose of radiation at each dose,CAL27 cells showed the highest and SCC15 showed the lowerst apoptosis rate.The RNA and protein expression levels of METTL3 in CAL27 were significantly lower than those of SCC15.m6A quantification showed that the methylation modification in CAL27 cells was lower than that in SCC15.The expression of METTL3 was increased in CAL27 and SCC15 cells after radiation treatment.Knockdown of METTL3 increaced the apoptosis rate and decreased the clonogenesity,overession of METTL3 the decreaced the ap optosis rate and increased the clonogenecity of the cells.Conclusion:Regulation of METTL3 can affect the radiotherapy sensitivity of OSCCs,METTL3 may become a new target for radiosensitization of OSCCs.

关 键 词：METTL3 甲基化 放疗敏感性 口腔鳞癌 

分 类 号：R739.8[医药卫生—肿瘤]