miRNA-325-3p通过下调细胞角蛋白13的表达降低鼻咽癌细胞CNE1的放疗敏感性  

miRNA-325-3p reduces the radio-sensitivity of nasopharyngeal carcinoma CNE1 cells by down-regulating the expression of cytokeratin 13

作　　者：万佳[1] 王欢[1] 王锦[1] 施明[1] 余宏[1] WAN Jia;WANG Huan;WANG Jin;SHI Ming;YU Hong(Department of Otorhinolaryngology Head and Neck Surgery,the Fourth Affiliated Hospital of Kunming Medical University,Kunming 650021,Yunnan,China)

机构地区：[1]昆明医科大学第四附属医院耳鼻咽喉头颈外科,云南昆明650021

出　　处：《中国肿瘤生物治疗杂志》2020年第5期496-500,共5页Chinese Journal of Cancer Biotherapy

基　　金：云南省科技厅-昆明医科大学应用基础研究联合专项基金资助[No.2018FE001(-173)]。

摘　　要：目的:探究miRNA-325-3p及其靶基因细胞角蛋白13(cytokeratin 13,CK13)对鼻咽癌细胞CNE1的放疗敏感性的影响。方法:通过miRBase、Targetscan及Microcosm三大数据库预测miRNA-325-3p的潜在靶基因,并通过双荧光素酶活性检测实验进行验证,qPCR检测不同放射剂量下鼻咽癌细胞CNE1中miRNA-325-3p及其靶基因的表达水平变化,通过克隆形成实验观察不同放射剂量下过表达miRNA-325-3p及敲低靶基因后CNE1细胞克隆形成率的变化,流式细胞术验证过表达miRNA-325-3p及敲低靶基因后CNE1在不同放射剂量下凋亡水平的变化,MTT法检测miRNA-325-3p过表达和CK13敲低组鼻咽癌细胞CNE1在不同放射剂量下的细胞存活率以验证其放疗敏感性的变化。结果:CK13确认为miRNA-325-3p的潜在靶基因,鼻咽癌细胞CNE1经放射处理后,miRNA-325-3p的表达水平显著升高、CK13的表达水平显著降低(均P<0.05)。miRNA-325-3p表达量上调和CK13基因沉默均显著提高CNE1细胞的存活率[miRNA上调时(:60.14±3.55)%vs(19.23±3.42)%,t=14.37、P<0.01;CK13沉默时:(76.15±5.13)%vs(28.53±3.68)%,t=13.06、P<0.01]和克隆形成率,降低了凋亡率[miRNA上调时:(27.95±2.67)%vs(51.68±3.47)%,t=9.39、P<0.01;CK13沉默时:(20.31±2.62)%vs(38.14±3.83)%,t=6.66、P<0.01]。结论:miRNA-325-3p能够通过下调靶基因CK13的表达降低鼻咽癌细胞CNE1对放疗的敏感性。Objective:To investigate the effect of miRNA-325-3 p and its target gene cytokeratin 13(CK13) on the radio-sensitivity of nasopharyngeal carcinoma cell line CNE1. Methods:The potential target gene of miRNA-325-3 p was predicted by three databases:miRBase, Targetscan and microcosm, and verified by Double luciferase activity assay. QPCR was used to detect the expression levels of miRNA-325-3 p and its target gene in nasopharyngeal carcinoma cell line CNE1 under different radiation doses;To verify the changes in radio-sensitivity of nasopharyngeal carcinoma cells, colony formation assay, Flow cytometery and MTT were used to observe the clone formation, apoptosis and cell viability of CNE1 cells after overexpression of miRNA-325-3 p and knockdown of CK13 under different radiation doses, respectively. Results:CK13 was confirmed as a potential target gene of miRNA-325-3 p. After radiotherapy, the expression level of miRNA-325-3 p in CNE1 cell was significantly increased, while the expression level of CK13 was decreased(all P<0.05);up-regulation of miRNA-325-3 p expression and silence of CK13 gene increased cell survival rate(upregulation of miRNA-325-3 p: [60.14±3.55]% vs [19.23±3.42]%, t=14.37, P<0.01;silence of CK13: [76.15±5.13]% vs [28.53±3.68]%, t=13.06, P<0.01)and colony formation rate, and decreased apoptosis rate(upregulation of miRNA-325-3 p: [27.95±2.67]% vs [51.68±3.47]%, t=9.39,P<0.01;silence of CK13: [20.31±2.62]% vs [38.14±3.83]%, t=6.66, P<0.01). Conclusion:miRNA-325-3 p can reduce the sensitivity of nasopharyngeal carcinoma cell line CNE1 to radiotherapy by down-regulating the target gene CK13.

关 键 词：miRNA-325-3p CK13 放疗敏感性 鼻咽癌 CNE1细胞 

分 类 号：R739.63[医药卫生—肿瘤] R730.2[医药卫生—临床医学]