塞来昔布对人高转移性鼻咽癌CNE-2Z细胞侵袭力的影响  

Influence of celecoxib on invasiveness of human high-metastatic nasopharyngeal carcinoma cell line CNE-2Z

作　　者：罗伟仁[1] 李丽霞[2] 黎思怡[3] 姜汉国[1] 陈小毅[1] 

机构地区：[1]广东医学院病理教研室,东莞523808 [2]广东医学院附属医院肿瘤中心 [3]广东医学院临床技能培训中心,东莞523808

出　　处：《中华耳鼻咽喉头颈外科杂志》2010年第11期941-945,共5页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：广东省自然科学基金项目（No.031963）;广东医学院青年基金项目（No.XQ0608）

摘　　要：目的 研究塞来昔布（Cox-2选择性抑制剂）对人鼻咽癌CNE-2Z细胞侵袭力的影响及其相关机制.方法 用四甲基偶氮唑蓝（MTT）法检测不同浓度塞来昔布对鼻咽癌CNE-2Z细胞增殖的影响.不同浓度塞来昔布作用鼻咽癌CNE-2Z细胞24 h后,Transwell小室检测细胞侵袭力的改变;免疫细胞化学检测肿瘤细胞中E钙黏素蛋白表达变化,反转录聚合酶链反应（RT-PCR）检测细胞中Cox-2和E钙黏素mRNA表达变化.结果 塞来昔布明显抑制CNE-2Z细胞存活率,呈明显的浓度依赖性,24 h后25、50、100 μmol/L塞来昔布组细胞存活率（-x±s,下同）分别为（94.75±1.34）%、（91.77±2.70）%、（64.54±1.20）%,48 h后细胞存活率分别为（88.41±1.28）%、（78.84±1.56）%、（52.46±2.25）%,50%抑制浓度（IC50）为100 μmol/L,同一时间点细胞存活率差异有统计学意义（F值分别为462.204和1328.306,P值均＜0.01）.0、25、50 μmol/L塞来昔布作用鼻咽癌CNE-2Z细胞24 h,侵袭实验显示穿膜细胞数（-x±s,下同）分别为（263.7±13.5）个、（185.3±8.7）个、（144.0±8.2）个,用药组穿膜细胞数明显减少,组间差异有统计学意义（F=102.089,P＜0.01）.免疫细胞化学结果显示,0、25、50μmol/L塞来昔布组E钙黏素阳性表达的CNE-2Z细胞数分别为（21.7±2.6）个、（28.7±2.4）个、（40.3±1.3）个,50 μmol/L组阳性细胞数明显增多（F=78.637,P＜0.01）.RT-PCR结果显示：25、50 μmol/L组Cox-2 mRNA的表达比对照组明显下降（t值分别为23.950和36.651,P值均＜0.01）;25、50 μmol/L塞来昔布组E钙黏素mRNA的表达明显高于对照组（t值分别为35.829和81. 497,P值均＜0.01）.结论 塞来昔布抑制鼻咽癌CNE-2Z细胞的侵袭能力,其机制可能与E钙黏素表达升高有关.Objective To investigate the effect and mechanism （a selective cyclooxygenase-2 inhibitor） on invasive ability of human nasopharyngeal carcinoma （NPC） line CNE-2Z. Methods The proliferation of NPC cells was examined by MTT assay. The invasive and migrating ability of NPC cells was detected with transwell chamber. E-cadherin protein expression was detected by immunocytochemistry and the expressions of Cox-2 and E-cadherin mRNA were analyzed by reverse transcription-polymerase chain reaction （RT-PCR）. Results MTT showed that celecoxib inhibited CNE-2Z proliferation in dose-dependent manner, the survival rate of cells treated with 25, 50, 100 μ mol/L celecoxib （-x±s） for 24 h was（94. 75 ±1.34）%, （91.77 ±2. 70）% , （64. 54 ± 1.20）%, respectively, and the survival rate of cells treated for 48 h was （ 88.41±1.28 ） %, （ 78. 84 ± 1.56 ） %, （ 52. 46 ± 2. 25 ） %, respectively, the concentration of 50% inhibition concentration of a substance （IC50） was 100 μmol/L, the difference was statistically significant between different concentration groups in the same time-point （ repectively, F were 462. 204 and 1328. 306, P 〈0. 01 ）. Treated with different concentrations of celecoxib（0, 25, 50 μmol/L） for 24, the cell numbers （-x±s） through PVPF by tumor invasion assay were （263.7 ± 13.5）, （185.3 ±8.7） and （144. 0 ± 8. 2）, the difference was statistically significant between the experimental and control group （F =102. 089, P 〈0. 01 ）. Immunocytochemistry showed that celecoxib significantly induced the increase of Ecadherin protein expression, also with a dose-dependence in 0 μmol/L, 25 μmol/L, 50 μmol/L group was （21.7 ±2. 6）, （28. 7 ±2. 4）, （40. 3 ± 1.3）, and 50 μmol/L group increased significantly （ F =78. 637,P 〈0. 01 ）. RT-PCR showed that celecoxib reduced the expression of Cox-2 mRNA expression in 25, 50 μmol/L group decreased significantly compared with the control group （respectively, t were 23. 950 and 36. 65

关 键 词：碘胺类 吡唑类 钙黏着糖蛋白类 鼻咽肿瘤 癌 鳞状细胞 肿瘤细胞 培养的 肿瘤浸润 

分 类 号：R686[医药卫生—骨科学]