乙醇诱导舌鳞状细胞癌上皮细胞间质转化  

Ethanol induces epithelial-mesenchymal transition in tongue squamous cell carcinoma

作　　者：马辉彬 王成[2] 蔡伟鑫[2] 侯劲松[2] 刘习强[2] 陈小华[2] 黄洪章[2] 

机构地区：[1]汕头大学医学院第一附属医院口腔外科,515041 [2]中山大学光华口腔医学院·附属口腔医院,广东省口腔医学重点实验室,广州510055

出　　处：《中华口腔医学研究杂志（电子版）》2016年第6期365-371,共7页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：国家自然科学基金(81272949、81202136、81572661);高等学校博士学科点专项科研基金(20120171120068);中央高校基本科研业务费专项资金(11ykpy47);广州市珠江科技新星专项(2014J2200045)

摘　　要：目的探讨乙醇对舌鳞状细胞癌（TSCC）细胞上皮间质转化的影响。方法乙醇诱导TSCC细胞株SCC9和SCC15,倒置显微镜下观察处理前后细胞形态及排列情况;Wound healing和Transwell实验检测乙醇处理前后细胞迁移和侵袭能力;Western blot和免疫荧光检测E-cadherin和Vimentin表达情况。数据分析采用SPSS 19.0统计软件,计数资料之间比较采用卡方检验,E-cadherin和Vimentin蛋白表达变化采用单因素方差分析。结果倒置显微镜下可见乙醇诱导后的SCC9和SCC15细胞由上皮细胞形态向间质细胞形态转化,细胞失去极性和紧密连接,由多边形、铺路石样转变为类似成纤维细胞样形态,细胞呈梭形,部分伸出伪足,细胞间隙增宽甚至丧失,排列较为杂乱、松散;Wound healing实验中,乙醇处理后的SCC9和SCC15细胞划痕愈合更快,迁移能力明显增强（χ^2SCC9,24 h=15.668,PSCC9,24 h=0.001;χ^2SCC15,24 h=4.703,PSCC15,24 h=0.028;χ^2SCC9,48 h=3.391,PSCC9,48 h=0.042;χ^2SCC15,48 h=2.386,PSCC15,48 h=0.136）;Transwell实验则证实乙醇处理后的SCC9和SCC15细胞的侵袭能力明显增强,SCC9相对侵袭率为384.24%（χ^2=26.917,P〈0.001）,SCC15相对侵袭率为428.49%（χ^2=32.880,P〈0.001）;Western blot和免疫荧光结果显示乙醇诱导后SCC9和SCC15细胞E-cadherin表达下调（F=131.533,PSCC9〈0.001,PSCC15=0.001）,Vimentin表达上调（F=355.388,PSCC9〈0.001,PSCC15〈0.001）。结论乙醇可诱导TSCC细胞发生上皮间质转化,促进其侵袭和迁移。Objective To evaluate the effect of ethanol on cell migration and motility in tongue squamous cell carcinoma（TSCC）cell lines and investigate its potential mechanisms. Methods Two TSCC cell lines SCC9 and SCC15 were cultured with DMEM/F12 medium and treated with or without ethanol. Cell morphology and growth patterns were observed under the light microscope. Wound healing and transwell assay were performed to determine the ability of migration and invasion. Western blot and immunofluorescence were used to detect the expressions of E-cadherin and Vimentin. Data were analyzed by Chi square test using SPSS 19.0 statistical software,and the protein level of E-cadherin and Vimentin were evaluated by One-Way ANOVA. Results SCC9 and SCC15 cells presented with cobblestone-like pattern and an apical-basal polarity,whereas,both of them demonstrated a dispersed,spindle-shaped morphology with migratory protrusions after treated with ethanol. In wounding healing assay,the cell motility of SCC9 and SCC15 cells was enhanced significantly after alcohol treatment（χSCC9,24 h^2= 15.668,PSCC9,24 h=0.001;χ^2SCC15,24 h=4.703,PSCC15,24 h=0.028;χ^2SCC9,48 h=3.391,PSCC9,48 h=0.042;χ^2SCC15,48 h=2.386,PSCC15,48 h=0.136）,while in transwell assay,the relative invasion rate was 384.24%（χ^2= 26.917,P〈0.001）in SCC9 compared to 428.49%（χ^2= 32.880,P〈0.001）in SCC15,which confirmed that invasion ability was significantly increased in SCC9 and SCC15 cells treated with ethanol. Moreover,E-cadherin expression was decreased（F = 131.533,PSCC9〈0.001,PSCC15= 0.001）,but Vimentin expression was increased（F =355.388,PSCC9〈0.001,PSCC15〈0.001）after administration of ethanol. Conclusion Ethanol could induce epithelial mesenchymal transition and promote migration and invasion in tongue squamous cell carcinoma.

关 键 词：舌 癌 鳞状细胞 乙醇 上皮细胞-间充质细胞转换 E-钙粘着糖蛋白 

分 类 号：R739.86[医药卫生—肿瘤]