姜黄素对肝癌耐药细胞HepG2/ADM的增殖和阿霉素耐药性的影响  

Effect of curcumin on proliferation and adriamycin-resistance of drug-resistant hepatocarcinoma cell HepG2/ADM

作　　者：曹聪[1] 黄桂柳 黄赞松[2] 胡高裕 邓志华[2] 李广志[2] 陆文权 钟秋红[4] CAO Cong;HUANG Gui-liu;HUANG Zan-song;HU Gao-yu;DENG Zhi-hua;LI Guang-zhi;LU Wen-quan;ZHONG Qiu-hong(Department of General Practice,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,China;Department of Gastroenterology,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,China;Department of Internal Medicine,the People’s Hospital of Napo County,Napo 533900,China;Department of Ultrasound,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,China)

机构地区：[1]右江民族医学院附属医院全科医学科,广西百色市533000 [2]右江民族医学院附属医院消化内科,广西百色市533000 [3]广西百色市那坡县人民医院内科,那坡县533900 [4]右江民族医学院附属医院超声科,广西百色市533000

出　　处：《广西医学》2020年第8期976-980,共5页Guangxi Medical Journal

基　　金：广西自然科学基金(2014GXNSFAA118143 );广西医药卫生科研课题(Z20170224 );广西研究生教育创新计划项目(YCSW2017214);广西科技基地与人才专项(广西肝胆疾病临床医学研究中心研究课题)(桂科AD17129025-11)。

摘　　要：目的分析姜黄素对肝癌耐药细胞HepG2/ADM的增殖和阿霉素耐药性的影响。方法 (1)将HepG2/ADM细胞分为不同浓度药物组(加入5μg/mL、10μg/mL、20μg/mL、40μg/mL、60μg/mL的姜黄素)、阴性对照组(接种细胞但不加药物)及空白对照组(不接种细胞,只加培养基),培养24、48及72 h后检测细胞增殖情况。(2)将HepG2细胞、HepG2/ADM细胞分为药物组、阴性对照组(接种细胞但不加药物)及空白对照组(不接种细胞,只加培养基)。HepG2细胞药物组加入不同浓度阿霉素(0.1μg/mL、0.2μg/mL、0.4μg/mL、0.8μg/mL、1.6μg/mL);HepG2/ADM细胞药物组分两个亚组,一组加入不同浓度阿霉素(1.5μg/mL、3μg/mL、6μg/mL、12μg/mL、24μg/mL),另一组加入5μg/mL姜黄素和不同浓度阿霉素(1.5μg/mL、3μg/mL、6μg/mL、12μg/mL、24μg/mL)。培养48 h后检测细胞增殖情况,计算姜黄素的逆转倍数以及HepG2/ADM细胞耐药指数。(3)将HepG2/ADM细胞分为对照组(只加培养基)、阿霉素组(14μg/mL阿霉素)、姜黄素组(5μg/mL姜黄素)、阿霉素与姜黄素联合组(14μg/mL阿霉素+5μg/mL姜黄素)。作用48 h后,检测各组磷酸化p38丝裂原活化蛋白激酶(p38MAPK)蛋白的表达。结果 (1)不同浓度的姜黄素均可抑制肝癌耐药细胞HepG2/ADM的增殖。姜黄素分别作用48、72 h后,HepG2/ADM细胞的增殖抑制率随着药物浓度的增加而增加(均P<0.05);在5、10、20、40μg/mL姜黄素作用下HepG2/ADM细胞的增殖抑制率随着作用时间的延长而增加(均P<0.05)。(2)HepG2/ADM细胞对阿霉素的耐药指数为6.81,对阿霉素呈中度耐药;5μg/mL姜黄素对HepG2/ADM细胞阿霉素耐药性的逆转倍数为1.49。(3)对照组磷酸化p38MAPK蛋白水平高于其他组(均P<0.05);姜黄素组、阿霉素组磷酸化p38MAPK蛋白水平均高于阿霉素与姜黄素联合组(均P<0.05),但姜黄素组与阿霉素组之间差异无统计学意义(P>0.05)。结论姜黄素不仅可体外抑制肝癌耐药细胞HepG2/ADM的�Objective To analyze the effect of curcumin on proliferation and adriamycin-resistance of drug-resistant hepatocarcinoma cell HepG2/ADM. Methods(1)HepG2/ADM cells were allocated to drug groups with various concentrations(added with curcumin of 5 μg/mL, 10 μg/mL,20 μg/mL, 40 μg/mL and 60 μg/mL), negative control group(cells were inoculated without drugs) or blank control group(added with medium without cells), and then the proliferation of the cells were determined after 24, 48 and 72 hours of culture.(2)HepG2 cells or HepG2/ADM cells were divided into drug group, negative control group(cells were inoculated without drugs) and blank control group(added with medium without cells). The HepG2 cells in the drug group were added with different concentrations of adriamycin(0.1 μg/mL, 0.2 μg/mL, 0.4 μg/mL, 0.8 μg/mL and 1.6 μg/mL);the HepG2/ADM cells in the drug group were divided into two subgroups, then a subgroup was added with different concentrations of adriamycin(1.5 μg/mL,3 μg/mL, 6 μg/mL, 12 μg/mL and 24 μg/mL) and the other subgroup with 5 μg/mL curcumin and different concentrations of adriamycin(1.5 μg/mL,3 μg/mL, 6 μg/mL, 12 μg/mL and 24 μg/mL). After 48 hours of culture,the cell proliferation was measured, and the reversal index of curcumin and resistance index of HepG2/ADM cells were calculated.(3)HepG2/ADM cells were allocated to control group(added with medium only), adriamycin group(treated by 14 μg/mL adriamycin), curcumin group( treated by 5 μg/mL curcumin) or adriamycin+curcumin group(treated by 14 μg/mL adriamycin and 5 μg/mL curcumin). After 48 hours of intervention, the expression of phosphorylated p38 mitogen-activated protein kinase(p38 MAPK) protein was measured in each group. Results(1) Curcumin at different concentrations could inhibit the proliferation of drug-resistant hepatocarcinoma cell HepG2/ADM. After 48 and 72 hours of intervention with curcumin, the inhibitory rate of HepG2/ADM cells rose with the increase of drug concentration(all P<0.05);with curcumin interve

关 键 词：肝癌 肝癌细胞HEPG2 肝癌耐药细胞HepG2/ADM 姜黄素 细胞增殖 耐药 阿霉素 p38丝裂原活化蛋白激酶 磷酸化 

分 类 号：R735.7[医药卫生—肿瘤]