氯喹上调死亡受体5表达增强Huh7细胞对肿瘤坏死因子相关凋亡诱导配体的敏感性

Expression of death receptor 5 upreguleted by chloroquine enhances the sensitivity of Hnh7 cells to tumor necrosis factor related apoptosis-inducing ligand

目的 探讨氯喹对肝细胞癌Huh7细胞死亡受体5（DR5）表达及肿瘤坏死因子相关凋亡诱导配体（TRAIL）诱导的细胞增殖和凋亡的影响.方法 Huh7细胞分为对照组（1∶1000二甲基亚砜）、TRAIL组（50μg/L）、氯喹组（10μmol/L）及TRAIL＋氯喹组（TRAIL 50 μg/L＋氯喹10 μmol/L）,四甲基偶氮唑盐（MTT）法检测各组细胞的增殖活性;免疫荧光检测各组细胞DR5蛋白的表达情况;4＇,6＇-二脒基-2-苯基吲哚（DAPI）核染色观察细胞凋亡,并用蛋白质印迹法检测细胞凋亡指标切割的聚腺苷酸二磷酸核糖转移酶（cleaved PARP）的表达情况.结果 TRAIL处理可降低Huh7细胞增殖活性,以对照组细胞活力为对照时,氯喹组、TRAIL组及TRAIL＋氯喹组细胞增殖抑制率分别为（89±8）％、（53±10）％及（27±7）％;与TRAIL组相比,TRAIL＋氯喹组细胞增殖活力下降（t=3.922,P=0.017）.氯喹组上调细胞DR5的表达,TRAIL＋氯喹组可活化细胞凋亡信号,TRAIL组和TRAIL＋氯喹组细胞的凋亡率分别为（10.0±2.3）％和（20.4±4.0）％,两组比较差异有统计学意义（t=3.894,P=0.018）.结论 氯喹可通过上调Huh7细胞DR5的表达,增强细胞对TRAIL作用的敏感性.

Objective To explore the effect of chloroquine on death receptor 5 （DR5） expression of hepatocellular carcinoma Huh7 cells and cell proliferation and apoptosis induced by tumor necrosis factor related apoptosis-inducing ligand （TRAIL）.Methods Huh7 cells were divided into four groups：the control group （1∶1 000 dimethyl sulfoxide）,TRAIL group （50 μg/L）,chloroquine group （10 μmol/L） and TRAIL ＋chloroquine group （TRAIL 50 μg/L ＋ chloroquine 10 μmol/L）.Thiazolyl blue tetrazolium bromide （MTT） assay was used to determine the proliferation activity of cells,immunofluorescence was used to detect the expression of DR5,4＇,6-diamidino-2-phenylindole （DAPI） staining was used to observe cell apoptosis and Western blot was used to detect the expression of cleaved poly ADP-ribose polymerase （PARP）.Results TRAIL treatment could decrease Huh7 cells proliferation activity;when compared with the cell viability in the control group,the cell proliferation inhibition rate of chloroquine group,TRAIL group and TRAIL＋ chloroquine group was （89±8） %,（53±10） % and （27±7） %,respectively;compared with TRAIL group alone,cell proliferation activity was decreased in TRAIL＋ chloroquine group （t =3.922,P =0.017）.The expression of DR5 was upregulated in chloroquine group,and the cell apoptosis signaling was activated in TRAIL ＋ chloroquine group.The cell apoptosis rate of TRAIL group and TRAIL ＋ chloroquine group was （10.0±2.3） % and （20.4±4.0） %,respectively,and there was a statistical difference （t =3.894,P =0.018）.Conclusion Chloroquine can enhance the cell chemosensitivity to TRAIL treatment by upregulating the expression of DR5 in Huh7 cells.