miR-182在口腔鳞癌血清外泌体中的表达及其临床意义  

The expression of miR-182 in serum exosomes of oral squamous cell carcinoma and its clinical significance

作　　者：邓威 孟颖 王晨星[1] 金齐尧 毛广艳 李怀奇[1] 叶金海[1] DENG Wei;MENG Ying;WANG Chenxing;JIN Qiyao;MAO Guangyan;LI Huaiqi;YE Jinhai(Jiangsu Key Laboratory of Oral Diseases,Department of Oral and Maxillofacial Surgery,the Affiliated Stomatological Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京医科大学口腔疾病研究江苏省重点实验室,南京医科大学附属口腔医院口腔颌面外科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2021年第4期509-515,共7页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81371123);江苏高校优势学科建设工程资助项目(2018-87);江苏省“科教强卫工程”医学重点人才项目(ZDRCA2016087);东南大学-南京医科大学合作项目(JX105GSP2017DN03)。

摘　　要：目的:探讨血清外泌体中miR-182的表达水平及其在口腔鳞状细胞癌浸润中的临床意义。方法:通过TCGA数据库与文献相关研究相结合,筛选出在头颈鳞癌组织及头颈鳞癌细胞株外泌体中均高表达的miRNA;分别运用ExoQuick试剂盒和超速离心法提取血清及HEK293细胞培养上清液中的外泌体,结合透射电子显微镜、纳米粒子追踪分析仪、Western blot实验鉴定外泌体;通过实时定量PCR方法检验筛选出的miRNA在两种口腔癌细胞系HN6、CAL27及口腔癌患者血清外泌体中的表达水平;通过共孵育的方法将miR-182 antagomir加载到HEK293细胞外泌体中,CCK-8实验检测该外泌体处理的CAL27细胞增殖能力的变化。结果:miR-182在头颈鳞癌组织、口腔鳞癌细胞株及头颈鳞癌细胞株外泌体中均显著高表达;口腔癌患者血清外泌体miR-182表达水平与对照组相比升高,但差异无统计学意义(P>0.05),肿瘤浸润深度>5 mm的患者血清外泌体内miR-182含量相对于正常对照组明显升高(P<0.05);加载miR-182 antagomir的外泌体处理CAL27细胞后,CAL27细胞的增殖能力显著下降(P<0.05)。结论:miR-182在口腔癌中高表达,血清外泌体miR-182的表达水平与口腔鳞状细胞癌的肿瘤浸润深度有关,加载miR-182 antagomir的外泌体可以显著抑制CAL27细胞的增殖。Objective:To explore the clinical significance of the expression level of miR-182 in serum exosomes in the infiltration of oral squamous cell carcinoma.Methods:Combined with the TCGA database and related research in the literature,miRNAs highly expressed in exosomes of head and neck squamous cell carcinoma tissues and cell lines were screened.Exosomes were extracted using ExoQuick kit and ultracentrifugationfrom serum and culture supernatant of HEK293 cells,and identified with transmission electron microscopy,nanoparticle tracking analyzer and Western blot;the expression levels of miR-182 in two oral cancer cell lines,HN6,CAL27,and serum exosome were determined by real-time quantitative RT-PCR.The proliferation of CAL27 cells treated with miR-182 antagomir loaded HEK293 cell exosomeswas detected by CCK-8 experiment.Results:miR-182 was highly expressed in the exosomes of head and neck squamous cell carcinoma tissues,oral squamous cell carcinoma cell lines and head and neck squamous cell carcinoma cell lines.The expression levels of serum exosome miR-182 in patients with oral cancer increased,compared with those in the control group.However,the differences were statistically insignificant(P>0.05).The content of miR-182 in the serum exosomes of patients with tumor infiltration depth>5 mm was significantly higher than that of the control group(P<0.05).After CAL27 cells were treated with exosomes loaded with miR-182 antagomir,the proliferation ability of CAL27 cells decreased significantly(P<0.05).Conclusion:miR-182 is highly expressed in oral cancer and the expression level of serum exosome miR-182 is related to the depth of tumor infiltration of oral squamous cell carcinoma.The exosomes loaded with miR-182 antagomir significantly inhibit the proliferation of CAL27 cells.

关 键 词：miR-182 口腔鳞状细胞癌 外泌体 

分 类 号：K739.8[历史地理—历史学]