6-姜酚在不同环境中对人肝癌细胞株杀伤和化疗增敏作用的研究  

Cell-killing and sensitization effect of 6-gingerol on human hepatoma carcinoma cell in chemotherapy in different environment

作　　者：蒲华清[1] 王秉翔[2] 杜爱玲[3] 李英杰[1] 张志勉[1] 

机构地区：[1]山东大学齐鲁医院健康体检中心,济南250012 [2]山东大学齐鲁医院脊柱外科,济南250012 [3]山东大学化学与化工学院

出　　处：《中华老年医学杂志》2014年第4期424-428,共5页Chinese Journal of Geriatrics

摘　　要：目的对比6-姜酚在正常和低氧、低糖模式下对于人肝癌细胞株（HepG-2）细胞的杀伤和化疗增敏作用。方法通过浓度为5、10、15、20、40、60mg／L阿霉素（多柔比星，ADM）和25、50、100、200μmol／L6-姜酚刺激培养至对数期的HepG-2细胞，采用CCK-8试剂盒检测HepCr2细胞增殖反应。通过流式细胞术结合细胞凋亡检测试剂盒双标染色法检测药物作用后细胞的凋亡情况。采用实时聚合酶链式反应（real—timePCR）检测bet-2、bax及birc-5mRNA的表达情况。结果6-姜酚、ADM作用于HepG-2细胞后，细胞生长受到抑制。两种模式下6-姜酚组、阿霉素组的抑制率随浓度的升高而增强，抑制率具有浓度依赖性。正常培养条件下对照组、6-姜酚组、ADM组及6-姜酚＋ADM组HepG-2细胞凋亡百分比分别为（7．98±0．76）％、（9．63±1．00）O、（12．70±2．13）％、（19．92±1．41）％；低氧低糖条件下对照组、6-姜酚组、ADM组及6-姜酚＋ADM组HepG-2细胞凋亡百分比分别为（13．92±2．02）％、（19．36±1．22）％、（27．87±0．99）％、（38．63±2．25）％，两种培养条件下各浓度组细胞凋亡百分比均较对照组增高，差异有统计学意义（t值分别为7．250、5．259、12．185、8．140、15．000、47．576，P值分别为0．019、0．034、0．007、0．015、0．004、0．000），两药联用组的凋亡数最高，且低氧、低糖各浓度组的凋亡数高于正常培养各浓度组。real—timePCR检测结果表明，正常培养条件下与对照组比较，实验组bcl-2、birc-5表达降低，bax表达无变化。低氧低糖条件下与对照组比较实验组bcl-2、birc-5表达降低，bax表达增高。在低氧低糖环境下bcl-2的表达较正常培养条件下的表达增高，而bax、birc-5表达减少，bcl-2／ban高于正常培养组。结论6-姜酚可能通过下调birc-5的表达，降低Survivin蛋白抑制肿瘤细胞的凋亡的能力对HepG-2细Objective To compare the cell-killing and sensitization effect of 6-gingerol on human hepatoma carcinoma （HepG-2） cell in normal mode versus hypoxia-hypoglycemia mode in chemotherapy. Methods The HepG-2 cells was cultured to logarithmic phase and treated with adriamyein doxorubicin hydrochloride （ADM） （5,10,15,20,40,60 mg/L） and 6-gingerol（25,50,100, 200 /~mol/L）in different concentrations. Then the cell counting kit （CCK-8） assay kit was used to determine the proliferation inhibition of HepG-2 cells. Cell apoptosis was detected by combining flow cytometry and AnnexinV-FITC PI double staining after treated with different drugs. The expressions of bcl-2, bax and birc-5 mRNA in HepG-2 cells was detected by real time fluorescence quantitative polymerase chain reaction（RT-PCR） assay. Results 6-gingerol and ADM had a certain degree of growth inhibition on HepG-2 cells. In two modes, the inhibition ratios of the 6-gingerol and ADM were both increased along with the increase of the concentration, which showed a dose-dependent manner. Flow cytometry analysis demonstrated that the apoptosis rate in the control group, 6-gingerolgroup, ADM group and the 6-gingerol4-ADM group in the normal mode was （7.98±0.76）%, （9.63 ± 1.00） %, （12.70 ±2.13） % and （ 19.92 ± 1.41 ） %respectively. The apoptosis rate in the control group, 6-gingerol group, ADM group and the 6-gingerol＋ADM group in the hypoxia-hypoglycemia mode was （13.92 ± 2.02）%, （19.36 ± 1.22）%, （27.87 ± 0.99）% and （38.63 ± 2.25）% respectively. It demonstrated that the apoptosis rate was increased in the experimental groups as compared to the control group under the two culture conditions（the normal mode and the hypoxia- hypoglycemia mode）（ t=7. 250, 5. 259, 12. 185, 8. 140, 15.000, 47. 576,respectively, all P〈.0.05, 0.01 or 0. 001）. The combination group had the highest number of apoptosis cells, and the number of apoptosis cells was higher in hypoxia-hypoglycemia group than in normal culture group.

关 键 词：癌  肝细胞 抗肿瘤联合化疗方案 细胞凋亡 细胞低氧 

分 类 号：R735.7[医药卫生—肿瘤]