c—fos在喉鳞状细胞癌耐药细胞株TU177／VCR中的表达及意义  

Expression and significance of c-fos in resistant cell line TU177/VCR of larynx squamous cell carcinoma

作　　者：李国栋[1] 胡晓玲[2] 邢建峰 师如意[4] 李欣[1] 李剑锋 李同丽[1] Department of Otorhinolaryngology, Shanxi Provincial People＇s Hospital Affiliated to Shanxi Medical University, Taiyuan 030012, China(Li GD, Li X, Li JF, Li TL ） ; Department of Pharmacology, Shanxi Medical University, Taiyuan 030001, China （ Hu XL ） ; Function Laboratory, Shanxi Medical University, Taiyuan 030001, China （ Xing JF） ; Department of Cell Biology and Genetics, Shanxi Medical University, Taiyuan 030001, China（ Shi RY)

机构地区：[1]山西医科大学附属山西省人民医院耳鼻咽喉头颈外科,太原030012 [2]山西医科大学基础医学院药理教研室,太原030001 [3]山西医科大学基础医学院机能实验室,太原030001 [4]山西医科大学基础医学院细胞生物学与遗传学教研室,太原030001

出　　处：《中华耳鼻咽喉头颈外科杂志》2018年第4期281-285,共5页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：国家自然科学基金项目（81602176）;山西省青年科学基金项目（201701D221249,201601D202107）;山西医科大学博士启动基金（03201006）,山西医科大学青年基金（057487）;山西医科大学331基础医学科技培植基金（201219）

摘　　要：目的研究原癌基因c—fos在喉鳞状细胞癌细胞TU177多药耐药形成中的作用。方法采用药物浓度梯度递增法建立稳定的喉鳞状细胞癌耐药细胞系TU177／VCR；采用RT—PCR及蛋白质印迹法检测TU177与TU177／VCR中c—fos基因和蛋白表达差异；构建靶向人c—fos基因的敲低和过表达质粒，分别转染TU177／VCR和TU177细胞；四甲基偶氮唑蓝（MTT）法检测在TU177／VCR细胞敲低c．fos和在TU177细胞过表达c-fos后对化疗药物的敏感性。组间比较采用t检验。结果TU177／VCR与TU177相比较，对于长春新碱（VCR）、紫杉醇（TAX）、顺铂（DDP）、5-氟尿嘧啶（5-FU）的耐药倍数分别为26．25、7．33、2．41、5．50，表明TU177／VCR耐药株构建成功；与TU177细胞相比，c-fos在TU177／VCR细胞中表达明显多，二者差异有统计学意义（P〈0．05）；TU177／VCR细胞敲低c-fos后细胞耐药性降低，TU177／VCR空载组半抑制浓度（IC20）值为（306．2±6．3）μmol／L，敲低组IC50值为（81．3±3．9）μmol／L，敲低组IC50是空载组Ic50的27％（P〈0．05）；在TUl77细胞过表达c-fos后细胞耐药性增加，TU177细胞空载组IC50值为（55．3±9．4）μmol／L，过表达组IC50值为（288．1±7．3）izmol／L，过表达组IC50是空载组IC50的5．21倍（P〈0．05）。结论c—fos对喉鳞状细胞癌TU177／VCR细胞的多药耐药具有重要作用，有可能成为喉癌治疗新的分子靶点。Objective To explore the effect of c-fos on multidrug resistance of laryngeal cancer TU177 ceils. Method Increasing drug concentration gradient is adopted to establish the stability of the laryngeal cancer drug resistance in cell line ; RT-PCR and Western blot were used to detect difference of the c-fos between TU177 and TU177/VCR ceils; plasmids with human c-fos knockdown or over expression were transfected into TU177/VCR and TU177 ceils respectively, and the effects of different treatment on cell proliferation were investigated with MTT. Results The drug resistance of TU177/VCR cells was 26. 25-fold in vincristine （VCR） ,7. 33-fold in Paclitaxel （TAX）, 2. 41 in cisplatin （ DDP）, and 5. 50 in 5-fluorouracil （5-FU）, comparing with TU177 （ P〈 0.05）. The TU177/VCR cells had significantly higher c-fos expression compared to TU177 cells （ P 〈 0. 05）. The results showed that the IC50 values of 5-FU for the NC group and c-fos shRNA group were （306. 2 ± 6. 3 ） p, mol/L and （81.3 ± 3.9 ） μmol/L, respectively, which was decreased by 73% in the c-fos shRNA group compared to that in the NC group （P 〈0.05）. Similarly, the results showed that the IC50 values for 5-FU were （55.3 ±9.4） μmol/L in NC group and （288.1 ±7.3） μmol/L in e-los WT group, which was increased 5.21-fold in c-fos WT ceils. Conclusion C-fos plays important role in multidrug resistance of larynx cancer cell TU177/VCR, and might become a new molecular target for laryngeal cancer treatment.

关 键 词：喉肿瘤 癌 鳞状细胞 抗药性 肿瘤 c—fos TU177 

分 类 号：R739.65[医药卫生—肿瘤]