还原型谷胱甘肽对多柔比星处理HepG2细胞株效果的影响  

Effects of reduced glutathione against doxorubicin in HepG2 cell line

作　　者：张懿[1] 倪鎏达[2] 周丰[2] 杨峻[2] 程明亮[1] 傅青春[2] 陈成伟[1] 吴银霞[1] 

机构地区：[1]贵阳贵州医科大学,550001 [2]解放军第八五医院南京军区肝病中心

出　　处：《肝脏》2015年第10期786-790,共5页Chinese Hepatology

基　　金：中国肝炎防治基金会天晴肝病研究基金(TQGB20120016)

摘　　要：目的研究还原型谷胱甘肽(GSH)对多柔比星(DOX)处理HepG2细胞株增殖及凋亡的影响;探索核转录因子(NF)-κB p65、Bcl-2在DOX单药及联合GSH诱导HepG2细胞凋亡后的表达及其意义。方法实验分4组,空白组:培养液,对照组:培养液+HepG2细胞+RPMl l640培养基,DOX组:培养液+HepG2细胞+DOX,DOX+GSH组:培养液+HepG2细胞+DOX+GSH。MTT法检测HepG2细胞生长,流式细胞仪检测细胞早期凋亡率,实时荧光定量PCR法检测NF-κB p65 mRNA的表达,Western印迹检测NF-κB p65及Bcl-2的表达。结果 (1)DOX组和DOX+GSH组作用细胞24 h、48 h、72 h均能显著抑制HepG2细胞增殖,DOX组抑制率显著高于DOX+GSH组(P<0.05),且抑制率具有时间和剂量依赖性。(2)对照组凋亡率为(0.733±0.153)%,DOX组凋亡率为(28.400±0.007)%,DOX+GSH组凋亡率为(15.500±0.006)%;DOX组、DOX+GSH组分别与对照组相比,差异有统计学意义(P<0.01);DOX组与DOX+GSH组相比,差异有统计学意义(P<0.01)。(3)两组在处理细胞24 h后,DOX+GSH组NF-κB p65mRNA表达显著高于DOX组(P<0.05)。(4)DOX组NF-κB p65、Bcl-2表达较对照组增高,DOX+GSH组表达较DOX组表达增高。结论 GSH与DOX联合使用可导致DOX化疗效果下降,其机制可能是通过进一步上调NF-κB p65和Bcl-2的表达实现的。临床上使用DOX化疗的肿瘤患者应避免同时使用GSH。Objective To study the effects of reduced glutathione （GSH） on doxorubicin （DOX） induced proliferation and apoptosis in HepG2, and to investigate the expression of NF-nB p65, Bcl-2 in apoptostic HepG2 induced by DOX monotherapy and DOX combination with GSH, respectively. Methods Four groups were devided, including blank group： RPMI 1640 medium only; control group： HepG2 cells were cultured in the same volume of medium; DOX group： cells were cultured in medium containing DOX~ DOX ＋ GSH group., cells were cultured in medium containing DOX and GSH. MTT assay was performed to probe proliferation of HepG2 cell. Flow cytometry was carried out to determine early apoptosis rate of HepG2 cells. Real-time polymerase chain reaction was administered to investigate NF-kB p65 mRNA expression. Western blotting was used to document protein expression of NF-kB p65 and Bcl-2. The experimental data was analyzed by SPSS version 19.0 software. Results （1） HepG2 cell proliferation was inhibited time- and dose-dependently in both DOX group and DOX ＋ GSH group at 24 h or 48 h or 72 h. Additionally, the proliferation rate was higher in DOX group than that in DOX ＋ GSH group（P〈0.05）. （2） The apoptosis rate of HepG2 was （0. 733 ± 0. 153） % in control group, （28. 400 ± 0. 007）% in DOX group, （ 15. 500 ± 0. 006）% in DOX ＋ GSH group, respectively, which showed statistically significant difference between the later two groups and control group （P〈0.01）, as well as between DOX group and DOX ＋ GSH group （P〈0.01）. （3） The mRNA expression of NF-kB p65 in DOX ＋ GSH group was higher than that in DOX group （P〈 0.05）. （4） The protein expression of NF-kB p65 and Bcl-2 in DOX＋ GSH group was higher than that in DOX group （P〈 0. 05）. Conclusion DOX and GSH combination therapy could attenuate chemotherapeutic effect though a potential mechanism by increasing expression of NF-kB p65 and Bcl-2. The combination of DOX and GSH in chemotherapy for cancer patients should be

关 键 词：还原型谷胱甘肽 多柔比星 人肝癌细胞株 HEPG2 

分 类 号：R735.7[医药卫生—肿瘤]