天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (1): 38-42.doi: 10.11958/20150010

• 细胞与分子生物学 • 上一篇    下一篇

STAT-3抑制剂WP1066增强顺铂对口腔鳞状细胞癌侵袭能力抑制作用的体外研究

孔令平1,刘爱芹2,周旋2,任玉3,黄媛媛1,刘速1,张仑4   

  1. 1. 天津医科大学肿瘤医院
    2. 天津市肿瘤医院
    3. 天津医科大学
    4. 天津医科大学附属肿瘤医院、天津市肿瘤防治重点实验室颌面耳鼻喉肿瘤科
  • 收稿日期:2015-07-06 修回日期:2015-08-03 出版日期:2016-01-15 发布日期:2016-01-15
  • 通讯作者: △通讯作者 E-mail： zhanglun@tjmuch.com E-mail:kong_lingping@sina.com
  • 作者简介:孔令平 （1988）， 女， 硕士在读， 主要从事头颈部鳞癌基础研究
  • 基金资助:
    基金项目： 国家自然科学基金资助项目 （81172573）

The STAT-3 inhibitor WP1066 promotes oral squamous cell carcinoma invasiveness by cisplatin in vitro

KONG Lingping1， LIU Aiqin1， ZHOU Xuan1， REN Yu2， HUANG Yuanyuan1， LIU Su1， ZHANG Lun1△   

  1. 1 Department of Maxillofacial & E.N.T （ear, nose, and throat）Oncology, Tianjin Medical University Cancer Institute and Hospital; National Clinical Research Center of Cancer; Key Laboratory of Cancer Prevention and Therapy,Tianjin 300060, China; 2 Tianjin Medical University
  • Received:2015-07-06 Revised:2015-08-03 Published:2016-01-15 Online:2016-01-15
  • Contact: △通讯作者 E-mail： zhanglun@tjmuch.com E-mail:kong_lingping@sina.com

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摘要：  目的 探讨信号转导及转录激活因子 3 （STAT-3） 调控 miR-21 增强化疗药物顺铂抑制人口腔鳞状细胞癌侵袭能力的作用。方法 实验共分 3 组： 二甲基亚砜组 （DMSO 组）、 顺铂组 （DDP 组）、 小分子抑制剂+顺铂组（WP1066+DDP 组）。实时定量 PCR 检测 miR-21 表达水平； Western blot 检测 STAT-3 及 p-STAT-3、 基质金属蛋白酶组织抑制因子 3 （TIMP-3）、 基质金属蛋白酶 2/9 （MMP-2/9） 的表达； 用 Matrigel 基质生长实验和 Transwell 体外侵袭实验检测肿瘤细胞生长形成球形克隆、 侵袭能力； 荧光素酶报告基因实验检测 miR-21 的表达水平。结果 WP1066+DDP 处理组的 STAT3/pSTAT-3 表达水平比 DDP 组低； WP1066+DDP 组 miR-21 表达比前两组低； 通过 Transwell 小室聚碳酸酯膜的细胞数 WP1066+DDP 组少于 DDP 组； WP1066+DDP 组细胞生长形成球的能力明显较 DDP 组减弱； TIMP-3 蛋白表达较前两组升高； MMP-2/9 表达水平较前两组明显下调。荧光素酶实验证明 WP1066+ DDP 组的荧光素酶活性明显低于 DDP 组和 DMSO 组。结论 通过抑制舌癌细胞中 STAT-3 活性， 下调 miR-21 表达， 可以增强化疗药物抑制口腔鳞状细胞癌的侵袭能力， 为使 WP1066 联合 DDP 化疗成为靶向治疗人口腔鳞状细胞癌提供了实验依据。

关键词: 癌,鳞状细胞, 口腔肿瘤, 肿瘤侵润, STAT3转录因子, 顺铂, miR-21

Abstract: Objective To investigate the effect of signal transducers and activators of transcription 3 （STAT-3） on sen⁃ sitizing oral squamous cell carcinoma to cis-dichlorodiamineplatinum via downregulating miRNA-21. Methods Tscca and Tca8113P160 human tongue squamous cell carcinoma cell lines were employed in this study. WP1066 was used to suppress STAT-3 signaling pathway. Cells were divided into three groups: dimethyl sulphoxide (DMSO) group, cis-dichlorodiamine⁃ platinum (DDP) group and WP1066+DDP group. Transcription level of miR-21 was assessed by real-time PCR, while the expression levels of STAT-3, p-STAT-3, tissue inhibitor of metalloproteinase-3 (TIMP-3) and matrix metalloproteinase-2/ 9 (MMP-2/9 ) were evaluated by Western blot assay. Matrigel matrix and transwell assay were used to determine cancer cell colony formation and invasive ability respectively. Expression level of miR-21 was examined by luciferase reporter gene as⁃ say. Results Expression levels of STAT-3, pSTAT-3 and miR-21 were significantly suppressed by WP1066 treatment. The diameters of culture colony in cells treated with WP1066 and DDP were smaller than those in control group. The number of tongue cancer cells that migrated through the transwell membrane in WP1066 and DDP treated group was less than that in control group. Additionally, MMP- 2/9 expression decreased while TIMP- 3 increased dramatically in both cell lines in WP1066 + DPP group compared to the other two groups. Conclusion Reduction of STAT-3 can sensitize oral squamous cell carcinoma to cis-dichlorodiamineplatinum via downregulating miR-21. Our study shows that DDP, in combination with WP1066, might be used as a potential target in the treatment of human oral squamous cell cancer

Key words: carcinoma, squamous cell, mouth neoplasms, neoplasm invasiveness, STAT3 transcription factor, cisplatin, miR-21