甲基莲心碱逆转肝癌HepG2/thermotolerance细胞对阿霉素耐受性的作用  

Effect of Neferine on Adriamycin-resistance of Thermotolerant Hepatocarcinoma Cell Line HepG2/thermotolerance

作　　者：艾小红[1] 唐小卿[2] 刘艳萍[1] 刘华清[3] 董琳[3] 

机构地区：[1]南华大学第一附属医院肿瘤科,湖南衡阳421001 [2]中南大学药学院,湖南长沙410078 [3]南华大学肿瘤研究所,湖南衡阳421001

出　　处：《癌症》2007年第4期357-360,共4页Chinese Journal of Cancer

基　　金：国家自然科学基金(No.30472040)~~

摘　　要：背景与目的:如何成功地逆转耐热癌细胞的多药耐药性(multidrug resistance,MDR)是当前肿瘤热疗的研究热点。本研究探讨耐热肝癌细胞能否对阿霉素(adriamycin,ADR)产生耐受性,以及甲基莲心碱(neferine,Nef)能否逆转耐热肝癌细胞的阿霉素耐药性。方法:MTT法检测肿瘤细胞存活率,PI染色流式细胞仪检测细胞凋亡率,间接免疫荧光流式细胞术检测bcl-2表达。结果:在43℃环境中培养24h后,耐热肝癌细胞HepG2/thermotolerance的细胞存活率和细胞凋亡率分别为(89.6±5.4)%和(13.6±5.4)%,而非耐热肝癌细胞HepG2的细胞存活率和细胞凋亡率分别为(23.9±3.6)%和(68.9±7.3)%。正常培养环境情况下(37℃),ADR对HepG2/thermotolerance细胞的IC50为(113.7±12.7)μmol/L,而对HepG2细胞的IC50为(10.5±2.3)μmol/L,耐药倍数达10.8倍。1、10、100μmol/LADR分别作用24h后,HepG2/thermotolerance细胞的凋亡率分别为(9.3±2.6)%、(17.8±7.3)%和(32.9±8.6)%,而HepG2细胞的凋亡率分别为(14.3±3.9)%、(38.9±6.8)%和(62.7±5.9)%。在37℃培养环境下,10、40μmol/LNef对HepG2细胞和HepG2/thermotolerance细胞无增殖抑制作用和凋亡诱导作用,但可使ADR对HepG2/thermotolerance细胞的IC50分别下降至(63.7±5.6)μmol/L和(16.8±2.8)μmol/L,逆转倍数分别为1.78和6.79,并可使10μmol/LADR对HepG2/thermotolerance细胞凋亡的诱导作用升高至(26.8±5.9)%和(34.9±8.7)%;HepG2/thermotolerance细胞较HepG2细胞高表达Bcl-2蛋白,而Nef能下调HepG2/thermotolerance细胞的Bcl-2表达。结论:HepG2/thermotolerance细胞对ADR可产生耐受性,Nef可逆转HepG2/thermotolerance细胞对ADR的耐受性,其机制可能与其下调HepG2/thermotolerance细胞Bcl-2蛋白表达有关。BACKGROUND ＆ OBJECTIVE： Nowadays, reversing the multidrug resistance （MDR） of thermotolerant carcinoma cells is a hot topic in tumor thermatology. This study was to investigate the adriamycin （ADR）-resistance of thermotolerant hepatocarcinoma cell line HepG2/thermotolerance and the effect of neferine （Nef） on the ADR-resistance of HepG2/thermotolerance cells. METHODS： Cell proliferation was measured by MTT assay. Cell apoptosis was detected by flow cytometry （FCM） with PI staining. The expression of Bcl-2 was measured by FCM using fluorescein isothiocyanate （FITC）-conjugated anti-bcl-2 antibodies. RESULTS： The proliferation rate and apoptosis rate of HepG2/thermotolerance cells cultured in 43℃ for 24 h were （89.6±5.4）% and （13.6±5.4）%, respectively; however, those of HepG2 cells were （23.9±3.6）% and （68.9±7.3）%, respectively. The 50% inhibition concentration （IC50） of ADR was 10.8 times higher for HepG2/thermotolerance cells than for HepG2 cells [（113.7±12.7） μmol/L vs. （10.5±2.3） μmol/L]. When treated with 1, 10, 100 μmol/L ADR at 37℃ for 24 h, the apoptosis rates of HepG2/thermotolerance cells were （9.3±2.6）%, （17.8±7.3）%, and （32.9±8.6）%, respectively, but those of HepG2 cells were （14.3±3.9）%, （38.9±6.8）%, and （62.7±5.9）%, respectively. In the presence of 10 and 40 μmol/L Nef, the IC50 of ADR for HepG2/thermotolerance cells was significantly decreased from （113.7±12.7） μmol/L to （63.7±5.6） μmol/L and （16.8±2.8） μmol/L, and the cell apoptosis induced by 10 μmol/L ADR was significantly increased from （17.8±4.3）% to （26.8±5.9）% and （34.9±8.7）%, respectively. Bcl-2 was overexpressed in HepG2/thermotolerance cells, whereas it was down-regulated when the cells were treated with 40 μmol/L Nef for 24 h. CONCLUSIONS： HepG2/thermotolerance cells are ADR-resistant. Nef may reverse the ADR-resistance of HepG2/thermotolerance cells by down-regulating Bcl-2 expression.

关 键 词：甲基莲心碱 耐热肝癌细胞 阿霉素 多药耐药性 BCL-2 

分 类 号：R735.7[医药卫生—肿瘤]