mTOR信号通路调控1,25二羟维生素D_3抑制喉癌细胞Hep-2细胞增殖的研究  

mTOR signaling pathway for regulating inhibition of 1,25 dihydroxyvitamin D_3 on Hep-2 cells proliferation in laryngeal carcinoma

作　　者：桂明才[1] 李兵[2] 戚思国 周长华[1] 

机构地区：[1]四川省达州市中心医院耳鼻咽喉头颈外科,635000 [2]重庆医科大学附属第一医院耳鼻咽喉科,重庆400016

出　　处：《国际检验医学杂志》2016年第15期2065-2067,共3页International Journal of Laboratory Medicine

基　　金：重庆市自然科学基金资助项目(cstc2013jcyjA10059)

摘　　要：目的研究1,25二羟维生素D3抑制喉癌细胞Hep-2细胞增殖作用以及对雷帕霉素靶蛋白(mTOR)信号通路的影响。方法用不同剂量1,25二羟维生素D3(10-8 mol/L、10-7 mol/L、10-6 mol/L)分别处理Hep-2细胞24、48、72h,四甲基偶氮唑蓝法(MTT)检测Hep-2细胞的增殖情况,并计算抑制率;采用流式细胞仪分析1,25二羟维生素D3对Hep-2细胞周期分布的影响,Western blot检测1,25二羟维生素D3对mTOR信号通路的影响。结果不同浓度1,25二羟维生素D3均可抑制Hep-2细胞增殖,改变细胞周期分布,使G0/G1期Hep-2细胞比例增高。1,25二羟维生素D3干预后Hep-2细胞TSC1、TSC2蛋白表达较对照组增高(P<0.01),Rheb蛋白表达明显降低;mTOR蛋白及其磷酸化水平表达与对照组比较均降低(P<0.01),mTOR蛋白磷酸化表达降低尤为明显(P<0.01);4EBP-1蛋白表达较对照组增高(P<0.01)。结论 1,25二羟维生素D3改变喉癌细胞Hep-2细胞周期分布,影响mTOR信号通路蛋白表达,从而抑制细胞增殖。Objective To explore the inhibition role of 1,25 dihydroxyvitamin D3on laryngeal cancer Hep-2cell proliferation and its influence on mTOR signal pathway.Methods Hep-2cells were treated with different concentrations of 1,25 dihydroxyvitamin D3（10-8,10-7,10-6 mol/L）for 24,48,72 hrespectively.The proliferation situation of Hep-2cells was detected by the MTT method and the inhibition rate was calculated.The effect of 1,25 dihydroxyvitamin D3on Hep-2cell cycle distribution was analyzed by flow cytometry.The influence of 1,25 dihydroxyvitamin D3on mTOR signaling pathway was detected by Western blot.ResultsDifferent concentrations of 1,25 dihydroxyvitamin D3could inhibit the proliferation of Hep-2cells,changed the cell cycle distribution and increased the proportion of Hep-2cells in G0/G1 phase.The expressions of TSC1 and TSC2protein after 1,25 dihydroxyvitamin D3intervention were increased compared with the control group（P〈0.01）,while the Rheb protein expression was significantly decreased（P〈0.01）：mTOR protein and phosphorylation level were significantly decreased compared with the control group（P〈0.01）,the decrease of mTOR protein phosphorylation was especially obvious（P〈0.01）;4EBP-1protein expression was increased compared with the control group（P〈0.01）.Conclusion 1,25-dihydroxyvitamin D3 alters the Hep-2cell cycle distribution,affects the protein expression of mTOR signaling pathway,thus inhibits the cell proliferation.

关 键 词：1 25二羟维生素D3 HEP-2细胞 增殖抑制 雷帕霉素靶蛋白 

分 类 号：R739.65[医药卫生—肿瘤]