摘要
目的:研究1,25-二羟维生素D3诱导人喉癌Hep-2细胞凋亡及其对PI3K/AKT/Bcl-2信号通路的影响。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24、48、72 h,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PI3K、AKT及其磷酸化、Bax和Bcl-2蛋白的表达水平。结果:MTT结果显示,1,25-二羟维生素D3可以抑制Hep-2细胞增殖(P<0.05),1,25-二羟维生素D3浓度为10-6 mol/L抑制率可达30.71%,在上述浓度内随着浓度增加和时间延长,抑制作用逐渐增强,呈时间-剂量依赖性。流式细胞术检测到10-8、10-7、10-6 mol/L 1,25-二羟维生素D3作用48 h后,Hep-2凋亡细胞比例显著增加,凋亡率分别为12.13%、14.05%、16.17%,高于空白对照组的6.82%(P<0.05)。Western blot检测结果显示1,25-二羟维生素D3处理48 h后,Hep-2细胞PI3K、AKT、p-AKT、Bcl-2蛋白表达降低,Bax蛋白表达升高(P<0.05)。结论:在一定浓度范围内,1,25-二羟维生素D3(10-6、10-7、10-8mol/L)能够抑制人喉癌Hep-2细胞增殖,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可通过影响PI3K/AKT/Bcl-2,从而诱导Hep-2细胞凋亡。
Objective：To study 1,25-dihydroxyvitamin D3 induced apoptosis in Hep-2 cells and its effect on PI3K/AKT/Bcl-2 signaling pathway. Methods：Hep-2 cells were processed by different concentrations of 1,25-dihydroxyvitamin D3 （10^4, 10-7 and 10^4 mol/L） for 24,48 and 72 h. The Hep-2 cells proliferation and inhibition rate were detected and calculated by methyl thiazolyl tetrazolium （MTr）. Apoptosis rate of Hep-2 cells was detected by flow cytometry. The expression levels of PI3K/AKT/Bel-2 were analyzed by Western blot. Results ：MTY results showed that 1,25-dihydroxyvitamin D3 can inhibit the proliferation of Hep-2 cells（P〈0.05）. 1,25-dihydroxyvitamin D3 concentration of 10^4 mol/L inhibition rate was up to 30.71% ,in a time-and dose-dependent manner. Flow cytometry showed 1,25-dihydroxyvitamin D3 with 10^-8, 10^-7 and 10^-4 mol/L significantly increased Hep-2 cell apoptosis rate after 48 h;the apoptosis rates were 12.13%, 14.05%, 16.17% respectively,higher than 6.82% of the normal control group（P〈0.05）. Western blot results showed that Bax protein expression increased and PI3K/AKT/p-AKT/Bel-2 protein expression decreased after 1,25- dihydroxyvitamin D3 treatment for 48 h. Conclusion：A certain range of concentration of 1,25-dihydroxyvitamin D3（ 10^-6, 10^-7 and 10^-8 mol/L） can inhibit proliferation of human laryngeal carcinoma Hep-2 Cells,in a time and dose dependent manner,probably through induced apoptosis. 1,25-dihydroxyvitamin D3 can up-regulate Bax protein expression and down-regulate PI3K/AKT/p-AKT/ Bel-2 protein expression to induce the apoptosis of Hep-2 cells.
出处
《重庆医科大学学报》
CSCD
北大核心
2017年第11期1422-1425,共4页
Journal of Chongqing Medical University
基金
重庆市自然科学基金资助项目(编号:cstc2013jcyj A10059)
