维罗非尼联合E3330对BRAFV600E突变甲状腺癌乳头状癌细胞系NIS水平影响  

Effect of vemurafenib combined with E3330 on NIS level of BRAFV600E mutant thyroid cancer cell line

作　　者：孙蓓 胡琳斐 王会娟 郑向前 SUN Bei;HU Lin-fei;WANG Hui-juan;ZHENG Xiang-qian(Cancer Hospital, Tianjin Medical University ,National Clinical Research Center for Cancer ,Tianjin Key Laboratory of Cancer Prevention and Therapy , Tianjin's Clinical Research Center for Cancer ,Tianjin 300060,P. R. China;Cancer Hospital, Tianjin Medical University ,National Clinical Research Center for Cancer ,Tianjin Key Laboratory of Cancer Prevention and Therapy , Tianjin's Clinical Research Center for Cancer ,Tianjin 300060,P. R. China)

机构地区：[1]天津医科大学肿瘤医院门诊办公室,国家肿瘤临床医学研究中心,天津市“肿瘤防治”重点实验室,天津市恶性肿瘤临床医学研究中心,天津300060 [2]天津医科大学肿瘤医院甲状腺颈部肿瘤科,国家肿瘤临床医学研究中心,天津市“肿瘤防治”重点实验室,天津市恶性肿瘤临床医学研究中心,天津300060

出　　处：《中华肿瘤防治杂志》2019年第15期1084-1089,共6页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的 放射性碘(radioactive iodine,RAI)是分化型甲状腺癌的主要治疗方法,然而许多患者会发生RAI耐药,可能与甲状腺癌钠碘转运体(sodium iodide symporter,NIS)的缺失或表达减少所引起的RAI摄入减少密切相关.本研究旨在探讨vemurafenib联合氧化还原因子-1(redox factor-1,Ref-1)抑制剂E3330对BRAFV600E突变型甲状腺乳头状癌细胞系NIS水平的影响,为临床放射性碘的有效治疗提供条件.方法 选取2014-01-01-2015-01-31天津医科大学肿瘤医院收治的124例甲状腺乳头状癌患者的肿瘤组织标本及其中8例患者正常组织.采用免疫组织化学染色法检测甲状腺乳头状癌和正常组织中Ref-1表达,同时对Ref-1表达情况进行评分并分析其与临床病理资料的关系;采用RT-qPCR、蛋白质印记法检测不同甲状腺乳头状癌细胞系中Ref-1表达水平;分别设置二甲基亚砜对照组(Ctrl组)、维罗非尼组(10μmol/L)、E3330组(50μmol/L)、联合组(10μmol/L维罗非尼+50μmol/L E3330),采用流式细胞学检测不同组别ROS含量变化及对分化指标NIS表达水平的影响.结果 BRAF突变(χ2=10.289,P=0.001)、有淋巴结转移(χ2=12.811,P<0.001)及TNMⅢ~Ⅳ期(χ2=4.216,P=0.04)与Ref-1的高表达呈正相关,Ref-1在甲状腺乳头状癌组织中mRNA表达(χ2=3.265,P=0.0056)高于正常组织,且在BRAF突变细胞系BCPAP中的蛋白表达高于其他细胞系及甲状腺正常细胞系.维罗非尼与E3330联用处理细胞后ROS水平出现降低(t=3.994,P=0.0162),同时分化指标NIS mRNA(t v=4.626,P v=0.0098;t c=9.694,P c=0.0006)及蛋白水平(t v=3.333,P v=0.029;t c=6.007,P c=0·0039)较对照组出现升高,且联合组NIS mRNA(t=2.836,P=0.0471)、蛋白水平(t=2.808,P=0.0484)的升高程度较维罗非尼组更高.NAC处理BCPAP细胞8 h清除ROS影响后,NIS mRNA(t=9.931,P=0.0006)和蛋白水平(t=5.388,P=0.0057)均出现升高,其差异有统计学意义.结论 维罗非尼联合抑制剂E3330可进一步提高其对甲状腺乳头状癌细胞OBJECTIVE Radioactive iodine(RAI) is the mainstay of treatment for differentiated thyroid carcinoma, nevertheless,RAI resistance occurs in many patients,which may be closely related to the reduction in RAI uptake caused by the loss or decreased expression of the thyroid cancer sodium iodine transporter( NIS). This study aimed to investigate the effect of vemurafenib combined with Ref-1 inhibitor E3330 on the level of NIS in BRAF^V600E mutant thyroid papillary carci noma cell line, and to provide conditions for clinical radioactive iodine effective therapy. METHODS Tissue samples from 124 patients with thyroid papillary carcinoma and fresh carcinoma tissues and normal tissues of 8 patients admitted to the Cancer Hospital of Tianjin Medical University from January 1,2014 to January 31,2015 were selected. The expressions of Ref-1 in papillary thyroid carcinoma and normal tissue were detected by immunohistochemical staining. The expression of Ref-1 was scored and analyzed with clinicopathological data. RT-qPCR and Western blot were used. The expression levels of Ref-1 in different papillary carcinoma cell lines were detected. Dimethyl sulfoxide control group (Ctrl group),vemurafenib group (10 μmol/L), E3330 group (50 μmol/L), and combination group (10 μmol/L vemurafenib + 50 μmol/L E3330) were used for flow cytometry,and detecting the changes of ROS content in different groups and the effect on the expression level of NIS. RESULTS The results in our study showed that BRAF mutation(X^2=10. 289,P = 0. 001),lymph node metastasis(X^2 =12. 811 ,P=0. 000) and poor TNM stage(X^2 =4. 216 ,P=0. 040) were positively correlated with high expression of Ref-1. APE1 expression in mRNA(t=3. 265,P=0. 005 6) of papillary thyroid carcinoma tissues was significantly higher than that in normal tissues. The protein expression in BRAF mutant cells BCPAP were significantly higher than those in other cell lines and normal thyroid cell lines. The levels of ROS mRNA and protein were significantly lower in cells treated with vemurafenib co

关 键 词：BRAF突变 氧化还原因子-1 甲状腺癌 钠碘转运体 

分 类 号：R736.1[医药卫生—肿瘤]