标题：肿瘤坏死因子对喉鳞状细胞癌细胞的增殖抑制作用

      作者：孙群 1，胡翔 2，胡福云 3    (深圳市宝安区石岩人民医院耳鼻喉科 1、普外科 2、外一科 3，广东 深圳 518108)

      卷次： 2017年28卷24期

      【摘要】 目的 探讨肿瘤坏死因子(TNF)对喉鳞状细胞癌细胞的增殖抑制作用。方法 选用人喉鳞癌Hep-2细胞株，分为对照组、20 pg/mL TNF-α组、50 pg/mL TNF-α组、100 pg/mL TNF-α组和 200 pg/mL TNF-α组，分别采用相应药物进行处理。采用MTT法检测细胞增殖抑制率，采用流式细胞术检测细胞凋亡率和细胞周期，采用免疫组织化学法检测细胞增殖细胞核抗原(PCNA)表达水平，采用Western blot法检测细胞周期蛋白依赖性激酶4 (CDK4)表达水平。结果 与对照组比较，各浓度TNF-α组Hep-2细胞增殖抑制率及凋亡率较高，且20 pg/mL TNF-α组<50 pg/mLTNF-α组<100 pg/mL TNF-α组和 200 pg/mL TNF-α组；与 24 h比较，相同浓度TNF-α作用 48 h后细胞增殖抑制率及凋亡率较高，差异有统计学意义(P<0.05)；与对照组比较，各浓度TNF-α组Hep-2细胞S期细胞比例较低，G2/M期细胞比例较高；且在20~100 pg/mL浓度范围内，TNF-α浓度越高，G2/M期细胞阻滞越严重(P<0.05)；与对照组比较，各浓度TNF-α组Hep-2细胞 PCNA表达水平及CDK4活性较低；且 20 pg/mL TNF-α组>50 pg/mL TNF-α组>100 pg/mL TNF-α组和200 pg/mL TNF-α组；与24 h比较，相同浓度TNF-α作用48 h后Hep-2细胞PCNA表达水平及CDK4活性较低，差异均有统计学意义(P<0.05)。结论 TNF对喉鳞状细胞癌Hep-2细胞增殖具有抑制作用，其能够使细胞阻滞于G2期，促进细胞凋亡，其作用可能与下调PCNA表达水平和CDK4活性有关。
      【关键词】 肿瘤坏死因子；喉鳞状细胞癌；细胞增殖；抑制；细胞周期；细胞增殖细胞核抗原
      【中图分类号】 R739.65 【文献标识码】 A 【文章编号】 1003—6350（2017）24—3961—05

Inhibitory effect of tumor necrosis factor on proliferation of laryngeal squamous cell carcinoma.
SUN Qun 1, HUXiang 2, HU Fu-yun 3. Shiyan People's Hospital of Shenzhen, Department of ENT 1, Department of General Surgery 2, FirstDepartment of Surgery 3, Shenzhen 518108, Guangdong, CHINA
【Abstract】 Objective To analyze the inhibitory effect of tumor necrosis factor (TNF) on proliferation of laryn-geal squamous cell carcinoma. Methods Human laryngeal squamous cell carcinoma Hep-2 cell were selected and di-vided into control group, 20 pg/mL TNF-αgroup, 50 pg/mL TNF-αgroup, 100 pg/mL TNF-αgroup, 200 pg/mL TNF-αgroup. Cells in each group were treated with the corresponding drugs. The cell proliferation inhibition rate was detectedby MTT method, and flow cytometry was used to detect apoptosis rate and cell cycle. The expression of proliferating cellnuclear antigen (PCNA) was detected by immunohistochemistry, and the expression levels of cyclin dependent kinase 4(CDK4) were detected by Western blot. Results Compared with control group, the proliferation inhibition rate and apop-·论 著·doi:10.3969/j.issn.1003-6350.2017.24.002基金项目：2014年广东省深圳市宝安区科技计划立项项目(编号：2014249)