YAP蛋白在口腔鳞癌中的表达及对肿瘤细胞迁移、侵袭的影响

中国口腔颌面外科杂志 ›› 2016, Vol. 14 ›› Issue (3): 203-208.

YAP蛋白在口腔鳞癌中的表达及对肿瘤细胞迁移、侵袭的影响

周榕¹, 石超吉¹, 韩永¹, 陶文杰¹, 李江², 孙树洋¹, 张雷¹, 张志愿¹

1.上海交通大学医学院附属第九人民医院·口腔医学院口腔颌面外科;
2.口腔病理科,上海市口腔医学重点实验室,上海 200011

收稿日期:2016-01-06 出版日期:2016-06-20 发布日期:2016-07-04
通讯作者: 张志愿,E-mail：zhzhy0502@163.com
作者简介:周榕（1989-）,男,硕士研究生,E-mail：santaneo@foxmail.com收集由上海交通大学医学院附属第九人民医院口腔颌面外科2007年2月—2008年5月间收治的经病理诊断为口腔鳞癌的患者68例,其中男45例,女23例。所有患者均未经过放疗或化疗。患者就诊年龄31~84岁,平均年龄55.5岁。所有患者均在签署书面知情同意书后,经手术切除肿瘤并进行肿瘤样本采集,之后经病理诊断进行确认,邻近正常上皮组织选取远离癌组织2cm以上的位置,以避免肿瘤细胞污染[18]。68例样本中,38例位于舌部,9例位于颊部,7例位于磨牙后区,7例位于口底,7例位于牙龈。根据WTOTNM以及肿瘤分期标准[19],由口腔病理科医师对肿瘤组织进行TNM及病理分级,并对患者其他相关临床信息进行采集。术后患者通过门诊复查及电话获得随访,随访时间为5~60个月。
基金资助:
国家自然科学基金（81001206）; 上海市科学技术委员会资助项目（08DZ2271100）

Yes-associated protein overexpression in oral squamous cell carcinoma and association with tumor cell migration and invasion

ZHOU Rong¹, SHI Chao-ji¹, HAN Yong¹, TAO Wen-jie¹, LI Jiang², SUN Shu-yang¹, ZHANG Lei¹, ZHANG Zhi-yuan¹

1.Department of Oral and Maxillofacial Surgery, 2. Department of Oral Pathology, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; Shanghai Key Laboratory of Stomatology. Shanghai 200011, China)

Received:2016-01-06 Online:2016-06-20 Published:2016-07-04

摘要/Abstract

摘要： 目的检测Yes相关蛋白 (Yes-associated protein,YAP)在口腔鳞状细胞癌(oral squamous cell carcinoma, OSCC)中的表达及临床意义,判断其是否与肿瘤细胞的侵袭和迁移特性相关。方法利用免疫组织化学染色检测68例原发口腔鳞癌患者组织标本中YAP的表达情况,分析其与各临床病理因素之间的关系;构建靶向YAP的shRNA以及存在特定位点突变的YAP过表达载体,转染含YAP蛋白的TB2T4和SCC23口腔鳞癌细胞;利用细胞划痕实验和细胞侵袭实验检测YAP干扰及过表达对肿瘤细胞迁移和侵袭能力的影响。采用SPSS18.0软件包对数据进行统计学处理。结果在原发口腔鳞癌组织中,YAP在细胞核和胞质的表达均显著高于正常口腔黏膜上皮组织(P<0.05)。YAP在细胞核的表达情况与患者颈淋巴结转移 (P<0.001)及5年总体生存率(P=0.008)显著相关。沉默YAP基因后,TB2T4和SCC23细胞的迁移和侵袭能力受到显著抑制 (P<0.05);过表达YAP基因后,肿瘤细胞的迁移和侵袭能力得到部分恢复（P<0.05)。结论YAP的表达与口腔鳞癌患者颈淋巴结转移及预后相关,YAP在口腔鳞癌细胞的侵袭、迁移中可能发挥重要作用。

关键词: 口腔鳞状细胞癌, Yes相关蛋白, 免疫组织化学染色, RNA干扰

Abstract: PURPOSE: To investigate the expression of YAP (Yes-associated protein) in oral squamous cell carcinoma (OSCC) and determine if YAP was associated with tumor cell invasion and migration. METHODS: Sixty-eight primary OSCC patients were enrolled in this study. YAP expression in tissue specimens was investigated by immunohistochemistry staining. YAP putback plasmid with specific mutation sites after YAP-shRNA interference was constructed by site-directed mutagenesis. Cell scratch and cell invasion assay were performed after YAP-siRNA interference or YAP putback again in TB2T4 and SCC23 cells. The data was analyzed using SPSS18.0 software package. RESULTS: Nuclear and cytoplasmic expression of YAP was increased in primary OSCC tissue samples (P<0.05). Nuclear expression of YAP was associated with cervical lymph node metastasis (P<0.001) and 5-year overall survival rate (P=0.008). YAP knockdown greatly suppressed tumor cell migration and cell invasion both in TB2T4 and SCC23 cells (Scramble vs YAP-shRNA, P<0.01), but these abilities were rescued subtotally when YAP was put back to the YAP-shRNA stable cells (Control vs YAP putback, P<0.01). CONCLUSION: YAP expression in OSCC was associated with cervical lymph node metastasis and YAP might play an essential role on the invasion ability of OSCC cells.

Key words: Oral squamous cell carcinoma, Yes-associated protein, immunohistochemistry, RNA interference

中图分类号:

R739.8

周榕, 石超吉, 韩永, 陶文杰, 李江, 孙树洋, 张雷, 张志愿. YAP蛋白在口腔鳞癌中的表达及对肿瘤细胞迁移、侵袭的影响[J]. 中国口腔颌面外科杂志, 2016, 14(3): 203-208.

ZHOU Rong, SHI Chao-ji, HAN Yong, TAO Wen-jie, LI Jiang, SUN Shu-yang, ZHANG Lei, ZHANG Zhi-yuan. Yes-associated protein overexpression in oral squamous cell carcinoma and association with tumor cell migration and invasion[J]. China J Oral Maxillofac Surg, 2016, 14(3): 203-208.

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