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  

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

作　　者：孔令平[1] 刘爱芹[1] 周旋[1] 任玉[2] 黄媛媛[1] 刘速[1] 张仑[1] 

机构地区：[1]天津医科大学肿瘤医院颌面耳鼻咽喉肿瘤科国家肿瘤临床医学研究中心天津市肿瘤防治重点实验室,300060 [2]天津医科大学

出　　处：《天津医药》2016年第1期38-42,共5页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(81172573)

摘　　要：目的探讨信号转导及转录激活因子3(STAT-3)调控mi R-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mi R-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mi R-21的表达水平。结果WP1066+DDP处理组的STAT3/p STAT-3表达水平比DDP组低;WP1066+DDP组mi R-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mi R-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。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 WP 1066 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 downregnlating 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.

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 

分 类 号：R739.8[医药卫生—肿瘤]