切换至 "中华医学电子期刊资源库"
高级检索
中华口腔医学研究杂志(电子版)

中华口腔医学研究杂志(电子版) ›› 2016, Vol. 10 ›› Issue (06) : 365 -371. doi: 10.3877/cma.j.issn.1674-1366.2016.06.001

所属专题： 文献；

基础研究

乙醇诱导舌鳞状细胞癌上皮细胞间质转化
马辉彬1, 王成2, 蔡伟鑫2, 侯劲松2, 刘习强2, 陈小华2, 黄洪章2,†()   
  1. 1. 515041　汕头大学医学院第一附属医院口腔外科
    2. 510055　广州，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
  • 收稿日期:2016-02-14 出版日期:2016-12-01
  • 通信作者: 黄洪章
  • 基金资助:
    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)

Ethanol induces epithelial-mesenchymal transition in tongue squamous cell carcinoma

Huibin Ma1, Cheng Wang2, Weixin Cai2, Jinsong Hou2, Xiqiang Liu2, Xiaohua Chen2, Hongzhang Huang2,†()   

  1. 1. Department of Oral Surgery, The First Affiliated Hospital of Shantou University Medical College, Shantou 515041, China
    2. Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China
  • Received:2016-02-14 Published:2016-12-01
  • Corresponding author: Hongzhang Huang
  • About author:
    Corresponding author: Huang Hongzhang, Email:
全文 ( ) 下载PDF ( ) 导出引用
引用本文：

马辉彬, 王成, 蔡伟鑫, 侯劲松, 刘习强, 陈小华, 黄洪章. 乙醇诱导舌鳞状细胞癌上皮细胞间质转化[J]. 中华口腔医学研究杂志(电子版), 2016, 10(06): 365-371.

Huibin Ma, Cheng Wang, Weixin Cai, Jinsong Hou, Xiqiang Liu, Xiaohua Chen, Hongzhang Huang. Ethanol induces epithelial-mesenchymal transition in tongue squamous cell carcinoma[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2016, 10(06): 365-371.

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（χ2SCC9，24 h= 15.668，PSCC9，24 h= 0.001；χ2SCC15，24 h= 4.703，PSCC15，24 h= 0.028；χ2SCC9，48 h= 3.391，PSCC9，48 h= 0.042；χ2SCC15，48 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%（χ2= 26.917，P<0.001），SCC15相对侵袭率为428.49%（χ2= 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。

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
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 (χ2SCC9, 24 h=15.668, PSCC9, 24 h= 0.001; χ2SCC15, 24 h=4.703, PSCC15, 24 h=0.028; χ2SCC9, 48 h=3.391, PSCC9, 48 h=0.042; χ2SCC15, 48 h = 2.386, PSCC15, 48 h= 0.136) , while in transwell assay, the relative invasion rate was 384.24% (χ2= 26.917, P<0.001) in SCC9 compared to 428.49% (χ2= 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.

Key words: Tongue, Carcinoma, squamous cell, Ethanol, Epithelial-mesenchymal transition, E-cadherin
图1 乙醇诱导前后SCC9、SCC15倒置显微镜下细胞形态学观察（× 40）
图2 乙醇诱导前后SCC9、SCC15细胞迁移能力检测（× 20）
图3 乙醇诱导前后SCC9、SCC15细胞侵袭能力检测
图4 免疫荧光检测E-cadherin和Vimentin表达（× 200）
图5 Western blot检测E-cadherin和Vimentin表达
[1]
Pöschl G, Seitz HK. Alcohol and cancer[J]. Alcohol Alcohol，2004，39(3):155-165.
[2]
Boffetta P, Hashibe M. Alcohol and cancer[J]. Lancet Oncol，2006，7(2):149-156.
[3]
Goldstein BY, Chang SC, Hashibe M，et al. Alcohol consumption and cancers of the oral cavity and pharynx from 1988 to 2009：an update[J]. Eur J Cancer Prev，2010，19(6):431-465.
[4]
CancelaMde C, Ramadas K, Fayette JM，et al. Alcohol intake and oral cavity cancer risk among men in a prospective study in Kerala，India[J]. Community Dent Oral Epidemiol，2009，37(4):342-349.
[5]
Ide R, Mizoue T, Fujino Y，et al. Cigarette smoking，alcohol drinking，and oral and pharyngeal cancer mortality in Japan[J]. Oral Diseases，2008，14(4):314-319.
[6]
Lissowska J, Pilarska A, Pilarski P，et al. Smoking，alcohol，diet，dentition and sexual practices in the epidemiology of oral cancer in Poland[J]. Eur J Cancer Prev，2003，12(1):25-33.
[7]
Wight AJ, Ogden GR. Possible mechanism by which alcohol may influence the development of oral cancer—a review[J]. Oral Oncol，1998，34(6):441-447.
[8]
Figuero Ruiz E, CarreteroPeláez MA, CereroLapiedra R，et al. Effects of the consumption of alcohol in the oral cavity：relationship with oral cancer[J]. Med Oral，2004，9(1):14-23.
[9]
Weinberg RA. Coevolution in the tumor microenvironment[J]. Nature Genet，2008，40(5):494-495.
[10]
Gupta GP, Massagué J. Cancer metastasis：building a framework[J]. Cell，2006，127(4):679-695.
[11]
Acloque H, Thiery JP, Nieto MA. The physiology and pathology of the EMT. Meeting on the epithelial-mesenchymal transition[J]. EMBO Rep，2008，9(4):322-326.
[12]
Savagner P. Leaving the neighborhood：molecular mechanisms involved during epithelial-mesenchymal transition[J]. Bioessays，2001，23(10):912-923.
[13]
Thiery JP. Epithelial-mesenchymal transitions in tumourprogression[J]. Nat Rev Cancer，2002，2(6):442-454.
[14]
Chamulitrat W, Schmidt R, Chunglok W，et al. Epithelium and fibroblast-like phenotypes derived from HPV16 E6/E7-immortalized human gingival keratinocytes following chronic ethanol treatment[J]. Eur J Cell Biol，2003，82(6):313-322.
[15]
Ke Z, Lin H, Fan Z，et al. MMP-2 mediates ethanol-induced invasion of mammary epithelial cells over-expressing ErbB2[J]. Int J Cancer，2006，119(1):8-16.
[16]
Forsyth CB, Tang YM, Shaikh M，et al. Alcohol stimulates activation of snail，epidermal growth factor receptor signaling，and biomarkers of epithelial-mesenchymal transition in colon and breast cancer cells[J]. Alcohol ClinExp Res，2010，34(1):19-31.
[17]
孙玉娟，孙正.乙醇与口腔癌临床流行病学的研究进展[J].国际口腔医学杂志，2009，36(2):218-220.
[18]
Huang WY, Winn DM, Brown LM，et al. Alcohol concentration and risk of oral cancer in puerto Rico[J]. Am J Epidemiol，2003，157(10):881-887.
[19]
Allen NE, Beral V, Casabonne D，et al. Moderate alcohol intake and cancer incidence in women[J]. J Natl Cancer Inst，2009，101(5):296-305.
[20]
Petti S, Scully C. Oral cancer：the association between nation-based alcohol-drinking profiles and oral cancer mortality[J]. Oral Oncol，2005，41(8):828-834.
[1] 张超, 童富云, 胡滢. 乳头、乳晕区原发性皮肤鳞状细胞癌一例[J]. 中华乳腺病杂志(电子版), 2020, 14(04): 258-259.
[2] 张文涵, 王成. 瘢痕鳞状细胞癌的研究进展[J]. 中华损伤与修复杂志(电子版), 2023, 18(01): 59-64.
[3] 岳志浩, 王晶, 闫子玉, 葛娜, 许向亮, 单小峰, 崔念晖. 牙槽外科相关舌神经损伤早期诊断及治疗中磁共振神经成像技术的应用[J]. 中华口腔医学研究杂志(电子版), 2023, 17(06): 413-417.
[4] 李晨曦, 谭小容, 魏巍, 李慕秋, 龚忠诚. 三级淋巴结构在口腔癌中的特征及意义[J]. 中华口腔医学研究杂志(电子版), 2023, 17(05): 315-321.
[5] 胡伟涛, 李星瀚, 刘琦, 孟贺, 马琳, 邓永强. 微小核糖核酸-21在口腔鳞状细胞癌中作为生物学标志物的研究进展[J]. 中华口腔医学研究杂志(电子版), 2022, 16(05): 328-332.
[6] 黄优, 侯伟, 潘永初, 王林. 两种功能矫治器在儿童开HE畸形治疗中的对比研究[J]. 中华口腔医学研究杂志(电子版), 2022, 16(05): 281-286.
[7] 周炼, 周航, 张东强, 徐海涛. 改良舌弓治疗下颌第一磨牙异位萌出的临床应用[J]. 中华口腔医学研究杂志(电子版), 2022, 16(02): 94-99.
[8] 李众, 姚小武, 陈仕生, 卢子正, 林敏校, 桂心伟. Toll样受体9和环氧合酶2在口腔鳞状细胞癌及癌旁组织中的表达意义[J]. 中华口腔医学研究杂志(电子版), 2021, 15(06): 333-340.
[9] 易晨, 黄子贤, 余东升. Hippo信号通路在口腔鳞状细胞癌中的研究进展[J]. 中华口腔医学研究杂志(电子版), 2020, 14(06): 396-400.
[10] 楚晨, 孙艳, 尚伟, 张晓春, 葛胜优. 安罗替尼的研究现状及其应用于晚期口腔鳞状细胞癌的治疗前景[J]. 中华口腔医学研究杂志(电子版), 2020, 14(05): 325-329.
[11] 徐嘉, 张子笠, 王玉龙. 原发性甲状腺鳞状细胞癌二例报道[J]. 中华普外科手术学杂志(电子版), 2020, 14(06): 647-647.
[12] 郭永坤, 单峤, 付旭东, 李培栋, 谢井伟, 周少龙, 刘婉清, 刘春颖. 显微血管减压术治疗原发性舌咽神经痛[J]. 中华神经创伤外科电子杂志, 2022, 08(05): 318-319.
[13] 陈星月, 陈新龙, 王逸平, 刘向新, 赵宏胜. 舌下微循环监测对脓毒症休克患者并发急性肾损伤的预测价值[J]. 中华重症医学电子杂志, 2022, 08(02): 147-152.
[14] 阿布地热合曼·吐尔孙尼牙孜, 石鑫, 郝玉军, 姜磊, 买买提江·卡斯木, 冯兆海, 裴祎楠. 显微血管减压术治疗舌咽神经痛疗效分析[J]. 中华脑科疾病与康复杂志(电子版), 2021, 11(02): 74-78.
[15] 王昊, 明倩文, 王斌, 卢太坤, 张海宁. 利奈唑胺致黑毛舌的临床诊断学特征[J]. 中华诊断学电子杂志, 2023, 11(04): 254-260.
阅读次数
全文


摘要

版权所有 中华医学会 中华医学电子音像出版社有限责任公司  京ICP备14006079号-1  网络出版服务许可证：(署)网出证(京)字第075号