槲皮素对热休克诱导人肝癌HepG2细胞HSP70和P-gp表达的影响  

Effect of Quercetin on Expressions of HSP70 and P-gp in Human Liver Heptocarcinoma HepG2 Cells Induced by Heat Shock

作　　者：唐武兵[1] 倪宏[1] 曾文铤[1] 马佩球[1] 朱科伦[1] 

机构地区：[1]广州医学院第一附属医院肝病研究室,广州510120

出　　处：《广州医学院学报》2002年第3期1-5,共5页Academic Journal of Guangzhou Medical College

基　　金：广东省中医药管理局科研基金 (10 10 4 5 ) ;广州医学院附一医院科研课题

摘　　要：目的 :研究热休克诱导人肝癌HepG2细胞热休克蛋白 70 (HSP70 )和P 糖蛋白 (P gp)随时间的表达规律 ,并研究应用槲皮素 (Qu)对二者表达的抑制效应。方法 :①使用恒温水浴法 4 2℃热休克HepG2细胞 90min ;②采用免疫细胞化学法、流式细胞术检测热休克前、热休克后 2、4、8、12、2 4h时HSP70和P gp的表达规律 ,以及热休克前 1h应用槲皮素在热后 4h二者的表达。结果 :① 4 2℃热休克诱导HepG2细胞 ,HSP70表达增多 ,“核内迁移” ,阳性细胞数升高 ,4h达到最高 (11.4 7% ) ,较对照组 (6 .6 4 % )升高近 2倍 (P <0 .0 0 5 ) ,2 4h后回落到低值 ,细胞核内表达减少 ;P gp阳性细胞数随HSP70升高后升高 ,12h达到最高值 (96 .31% ) ,较对照组 (33.95 % )升高近 3倍 (P<0 .0 0 5 ) ,2 4h后回落到低值 ,同时 ,细胞内低水平的HSP70表达在细胞质中 ,少部分在细胞核内。②热休克前应用槲皮素能有效抑制热休克诱导HSP70和P gp的过量表达 ,槲皮素并能有效抑制二者内在表达 ,以 10 0～ 2 0 0μmol/LQu作用明显 (P <0 .0 0 5 ) ,呈浓度剂量效应。结论 :①热休克可能诱导HepG2细胞HSP70和P gp的过量表达 ,P gp与HSP70具有相关关系 ;②槲皮素可能在转录水平上抑制HepG2细胞HSP70和PObjective:To study the expressions of heat shock protein 70 (HSP70) and P glycoprotein (P gp) in human heptocarcinoma line HepG2 cells in vitro after being induced by heat shock;and to study the inhibition effect of quercetin (Qu) on them. Methods 1. HepG2 cells were shocked being bathed in water at 42 ℃ stably for 90 min. 2. By the use of immunocytochemistry and flow cytometry (FCM),the expressions of HSP70 and P gp were detected at 0 hour before and 2,4,8,12,24 hours after 42 ℃ heat shock;Furthermore,they were detected at 4 h after heat shock when HepG2 cells were pretreated by quercetin (Qu) for an hour before 42 ℃ heat shock. Results 1. HSP70 overexpressed in cells and ”moved” from cytoplasm to nuclei and nucleolus after HepG2 cells were shocked at 42 ℃.The percentage of HSP70 positive cells came to a climax (11.47%) at 4 h and increased about 2 folds compared with that of the control group (6.64%)( P <0.005). At 24 h after heat shock,it declined to lower than that of the control group. At the same time,lower HSP70 expressed in cytoplasm and only a little were located in nuclei. The percentage of cells with P gp positive went up following HSP70 after heat shock. It came to a climax (96.31%) at 12 h. And it increased about 3 folds compared with that of the control group (33.95%)( P <0.005). 2. Not only the overexpressions of HSP70 and P gp induced by heat shock but also the innate expressions of the both could be inhibited effectively by Qu in a dose dependent manner,most obviously at a Qu level of 100～200 μmol/L( P <0.005). Conclusions 1. Heat shock is possible to induce the overexpressions of HSP70 and P gp in HepG2 cells. P gp possibly has a correlation with HSP70. 2. Qu possibly inhibits the productions of HSP70 and P gp in HepG2 cells at mRNA in a dose dependent manner.

关 键 词：槲皮素 肝肿瘤 热休克 热休克蛋白-70 P-糖蛋白 免疫组化 流式细胞术 

分 类 号：R735.7[医药卫生—肿瘤]