白藜芦醇对肾细胞癌ACHN细胞增殖与凋亡的影响  

Effect of resveratrol on proliferation and apoptosis of renal cell carcinoma ACHN cells

作　　者：邓廷志 李富林[1] 陈烈钳 黄栋强[1] 陈景宇[1] DENG Tingzhi;LI Fulin;CHEN Lieqian;HUANG Dongqiang;CHEN Jingyu(Department of Urology,the First Hospital of Huizhou,Huizhou 516000,China)

机构地区：[1]惠州市第一人民医院泌尿外科

出　　处：《中华实用诊断与治疗杂志》2019年第9期865-868,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：广东省医学科研基金(A2018235)

摘　　要：目的探讨白藜芦醇对肾细胞癌ACHN细胞增殖及凋亡的影响及可能机制。方法对数生长期ACHN细胞随机分为对照组(100μmol/L培养液)、25μmol/L白藜芦醇组(100μmol/L培养液+25μmol/L白藜芦醇)、50μmol/L白藜芦醇组(100μmol/L培养液+50μmol/L白藜芦醇)、100μmol/L白藜芦醇组(100μmol/L培养液+100μmol/L白藜芦醇)、200μmol/L白藜芦醇组(100μmol/L培养液+200μmol/L白藜芦醇)。培养12、24、48h,采用CCK-8法检测细胞增殖活力;培养24h,采用流式细胞仪检测细胞凋亡率,采用荧光探针法检测ACHN细胞氧化型二氯荧光素(dichlorofluorecin,DCF)荧光强度,采用Western blot法检测B淋巴细胞瘤-2基因(B-cell lymphom-2,Bcl-2)、caspase-3蛋白表达。结果对照组及25、50、100、200μmol/L白藜芦醇组培养12h(0.71±0.01、0.63±0.01、0.56±0.03、0.48±0.02、0.39±0.02)、24h(1.19±0.04、1.00±0.02、0.79±0.03、0.63±0.03、0.41±0.06)、48h(1.62±0.02、1.15±0.03、0.79±0.04、0.62±0.04、0.41±0.08)细胞增殖活力吸光度值依次降低(P<0.05),培养24h细胞凋亡率[(7.73±0.87)%、(12.64±1.13)%、(29.43±1.25)%、(37.73±1.24)%、(48.56±1.78)%]依次升高(P<0.05);培养24h,对照组及25、50、100、200μmol/L白藜芦醇组细胞DCF荧光强度[(5.40±0.70)、(14.97±1.33)、(17.03±0.80)、(19.67±1.16)、(31.50±2.28)u/(s·mg)]及caspase-3相对表达量(0.24±0.02、0.58±0.02、1.19±0.05、1.41±0.05、1.74±0.06)依次升高,Bcl-2相对表达量(1.67±0.10、1.11±0.16、0.94±0.11、0.67±0.09、0.53±0.02)依次降低(P<0.05)。结论白藜芦醇可诱导肾细胞癌ACHN细胞内活性氧累积,上调caspase-3表达,下调Bcl-2表达,进而抑制细胞增殖,促进细胞凋亡,且呈剂量依赖性。Objective To investigate the effect of resveratrol(RES)on the proliferation and apoptosis of renal cell carcinoma ACHN cells and its possible mechanism.Methods The ACHN cells in logarithmic growth phase were randomly cultured with 100μmol/L medium(control group),100μmol/L medium+25μmol/L RES(25μmol/L RES group),100μmol/L medium+50μmol/L RES(50μmol/L RES group),100μmol/L medium+100μmol/L RES(100μmol/L RES group),and 100μmol/L medium+200μmol/L RES(200μmol/L RES group),respectively.The proliferation activity was detected by CCK-8after 12,24and 48h,the apoptotic rate was detected by flow cytometry after 24h,the fluorescence intensity of oxidized dichlorofluorecin(DCF)in ACHN cells was measured by fluorescent probe method,and the expressions of Bcl-2and caspase-3proteins were detected by Western blot.Results The optical density(OD)of ACHN cells decreased gradually in turn in control group and 25,50,100and 200μmol/L RES groups after 12hculture(0.71±0.01,0.63±0.01,0.56±0.03,0.48±0.02,0.39±0.02),24hculture(1.19±0.04,1.00±0.02,0.79±0.03,0.63±0.03,0.41±0.06)and 48h(1.62±0.02,1.15±0.03,0.79±0.04,0.62±0.04,0.41±0.08)(P<0.05),while the apoptosis rate after 24hculture increased gradually in turn((7.73±0.87)%,(12.64±1.13)%,(29.43±1.25)%,(37.73±1.24)%,(48.56±1.78)%)(P<0.05).DCF fluorescence intensity of ACHN cells((5.40±0.70),(14.97±1.33),(17.03±0.80),(19.67±1.16),(31.50±2.28)u/(s·mg))and the relative expression level of caspase-3(0.24±0.02,0.58±0.02,1.19±0.05,1.41±0.05,1.74±0.06)increased gradually in turn in control group and 25,50,100and 200μmol/L RES groups,while the relative expression of Bcl-2(1.67±0.10,1.11±0.16,0.94±0.11,0.67±0.09,0.53±0.02)decreased gradually in turn(P<0.05).Conclusion RES can induce reactive oxygen species accumulation in ACHN cells,up-regulate the expression of caspase-3and down-regulate Bcl-2so as to inhibit cell proliferation and promote cell apoptosis in a dose-dependent manner.

关 键 词：肾细胞癌 白藜芦醇 ACHN细胞 增殖 凋亡 

分 类 号：R28[医药卫生—中药学]