1,25二羟维生素D3诱导喉癌细胞Hep-2细胞凋亡及其对Bax/Bcl-2蛋白表达水平的影响

Mechanisms of human laryngeal carcinoma Hep-2 cell proliferation inhibition and apoptosis induction by 1,25-dihydroxyvitamin D3

目的研究1,25二羟维生素D3抑制人喉癌Hep-2细胞增殖作用,诱导细胞凋亡及其凋亡相关机制。方法用不同剂量(10^(-8)、10^(-7)、10^(-6) mol/L)1,25二羟维生素D3处理Hep-2细胞24h、48h、72h,四甲基偶氮唑蓝法(MTT)检测Hep-2细胞的增殖情况,计算抑制率。流式细胞术检测Hep-2细胞的凋亡率。Western blot检测用药前后细胞中Bax和Bcl-2蛋白的表达水平。结果 1,25二羟维生素D3可以抑制Hep-2细胞增殖(P<0.05),最高抑制率可达30.71%,在上述浓度内随着浓度增加、时间延长抑制作用逐渐增强,呈时间-剂量依赖性。10-7、10-6 mol/L 1,25二羟维生素D3作用48h后,Hep-2凋亡细胞比例显著增加,凋亡率高于空白对照组(P<0.05)。1,25二羟维生素D3处理48h后,Hep-2细胞Bax蛋白表达上升,Bcl-2蛋白表达水平下降。结论在一定浓度范围内1,25二羟维生素D3能够抑制人喉癌Hep-2细胞的增殖,呈时间-剂量依赖性,其抑制作用与诱导细胞凋亡有关;1,25二羟维生素D3可通过上调Bax蛋白表达、下调Bcl-2蛋白表达诱导Hep-2细胞凋亡。

Objective To investigate the mechanisms of human laryngeal carcinoma Hep-2cell proliferation inhibition and apoptosis induction by 1,25-dihydroxyvitamin D3.Methods Hep-2cells were processed by different concentration of 1,25 dihydroxyvitamin D3（10 -8 and 10 -7 and 10 -6 mol/L）for 24 hrs,48hrs and 72 hrs.The Hep-2cells proliferation and inhibition rate were detected and calculated by methyl thiazolyl tetrazolium（MTT）.Apoptosis rate of Hep-2cells was detected by flow cytometry.The expression levels of Bax and Bcl-2were analyzed by western blot（WB）.Results MTT results were shown that 1,25-dihydroxyvitamin D3 can inhibit the proliferation of Hep-2cells（P〈0.05）.The highest inhibition rate reached 30.71%,in a time-and dose-dependent manner.Flow cytometry showed 1,25 dihydroxyvitamin D3with 10-7,10-6 mol/L significantly increased Hep-2cell apoptosis rate after 48hrs（P〈0.05）.West blot（WB）results were shown that Bax protein expression increased and Bcl-2protein expression decreased after 1,25-dihydroxyvitamin D3 treatment for 48 hrs.Conclusion A certain range of concentration of 1,25-dihydroxyvitamin D3（10-6 to 10-7 and 10-8 mol/L）can inhibit proliferation of human laryngeal carcinoma Hep-2Cells,in a time and dose dependent manner,probably through induced apoptosis.1,25-dihydroxyvitamin D3 can up-regulate Bax protein expression and down-regulate Bcl-2protein expression to induce the apoptosis of Hep-2cells.