硫利达嗪通过内质网应激介导DR5表达上调增敏TRAIL对肺癌PC9细胞的促凋亡效应

发布时间：2018-01-01 03:18

本文关键词：硫利达嗪通过内质网应激介导DR5表达上调增敏TRAIL对肺癌PC9细胞的促凋亡效应　出处：《中国肺癌杂志》2017年02期 　论文类型：期刊论文

【摘要】：背景与目的肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor-related apoptosis-inducting ligand,TRAIL)可诱导肿瘤细胞发生凋亡,然而相当数量的肿瘤细胞可耐受TRAIL诱导的凋亡而得以存活。本实验观察硫利达嗪(thioridazine,THZ)通过诱导内质网应激(endoplasmic reticulum stress,ER stress)介导的死亡受体5(death receptor 5,DR5)表达上调,继而增敏TRAIL对肺腺癌细胞PC9的生长抑制及凋亡诱导效应,探讨其机制。方法不同浓度硫利达嗪及TRAIL单独或联合处理PC9细胞,MTT法检测细胞活性变化,流式细胞术检测细胞表面DR5表达及细胞凋亡率,Western blotting检测内质网应激相关蛋白GRP78(glucose regulated protein 78)、C/EBP环磷酸腺苷反应元件结合转录因子同源蛋白(C/EBP homologous protein,CHOP)、p-PERK(PKR-like ER kinase)、p-e IF2α(eukaryotic initiation factor-2α,e IF2α)、ATF4(activating transcription factor 4,ATF4)及凋亡相关蛋白Caspase-3、Caspase-9、Caspase-8、PARP、DR5表达变化。结果硫利达嗪对PC9细胞的增殖抑制效应呈浓度依赖性(P0.05),硫利达嗪可增加TRAIL对PC9细胞的抑制作用及凋亡诱导作用且可上调PC9细胞表面DR5表达水平,流式细胞术结果显示:TRAIL联合硫利达嗪组细胞凋亡率较单独TRAIL组显著增加(P0.05),Western blotting结果显示:TRAIL联合硫利达嗪组细胞Cleaved-caspase-8、Cleaved-PARP、DR5表达水平较单独TRAIL组明显上调。DR5表达上调及促凋亡效应是通过诱导内质网应激发生,并伴随着GRP78及CHOP表达上调发生的,且该效应可被4-苯基丁酸(4-phenylbutyric acid,4-PBA)可抑制(P0.05)。结论硫利达嗪增敏TRAIL对PC9细胞的增殖抑制效应显著,其机制可能与硫利达嗪内质网应激介导的DR5上调有关。
[Abstract]:Background and objective tumor necrosis factor-related apoptosis-inducing ligands. Tumor necrosis factor-related apoptosis-inducting ligand. Trail can induce apoptosis of tumor cells. However, a considerable number of tumor cells could survive the apoptosis induced by TRAIL. THZ induced endoplasmic reticulum stress by endoplasmic reticulum stress. The expression of ER stress-mediated death receptor 5 death receptor 5 DR5 was up-regulated. Then the growth inhibition and apoptosis induction of lung adenocarcinoma cell line PC9 were investigated by sensitizing TRAIL. Methods PC9 cells were treated with different concentrations of talidazide and TRAIL alone or in combination. MTT assay was used to detect the changes of cell activity, and flow cytometry was used to detect the expression of DR5 and the rate of apoptosis. The endoplasmic reticulum stress-related protein (GRP78(glucose regulated protein 78) was detected by Western blotting. C / EBP homologous protein (CHOP5), a transcription factor homologous protein, is bound by the C / EBP cyclic adenosine monophosphate response element. P-PERKU PKR-like ER kinase factor-2 伪 -e IF2 伪 eukaryotic initiation factor-2 伪. E IF2 伪 ATF4activation transcription factor 4 ATF4) and apoptosis-related protein Caspase-3. Results the inhibitory effect of thiridamine on the proliferation of PC9 cells was in a concentration dependent manner (P 0.05). Tiridazide could increase the inhibitory effect and apoptosis induction of TRAIL on PC9 cells and up-regulate the expression of DR5 on the surface of PC9 cells. The results of flow cytometry showed that the apoptosis rate of TRAIL group was significantly higher than that of TRAIL group (P 0.05). The results of Western blotting showed that the cell line Cleaved-caspase-8 was Cleaved-PARP. Compared with TRAIL group, the expression of DR5 increased significantly. DR5 expression and apoptosis were induced by endoplasmic reticulum stress, accompanied by the up-regulation of GRP78 and CHOP expression. The effect can be obtained by 4-phenylbutyric acid. Conclusion tiridamine sensitized TRAIL can inhibit the proliferation of PC9 cells. The mechanism may be related to the upregulation of DR5 mediated by endoplasmic reticulum stress.
【作者单位】：安徽医科大学第三附属医院(合肥市第一人民医院)肿瘤科;合肥市滨湖医院中心实验室;
【基金】：国家重点外国专家项目(No.20163400008) 安徽省重点外国专家项目(No.20153400062)资助~~
【分类号】：R734.2
【正文快照】： 肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor-related apoptosis-inducting ligand,TRAIL)属于TNF超家族,可诱导肿瘤细胞发生凋亡。TRAIL可通过与其受体TRAIL-R1(death receptor 4,DR4)、TRAIL-R2(death receptor 5,DR5)结合促使靶细胞发生凋亡。然而,相当数量的肿瘤

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