摘要
目的:观察刺梨提取物(extract from Rosa roxburghiiTratt,CL)联合5-氟尿嘧啶(5-florouracil,5-FU)抗人肝癌细胞SMMC-7721的作用。方法:采用MTT法检测和生长曲线绘制分析CL联合5-FU对SMMC-7721细胞生长的影响;通过检测硝基四氮唑蓝(nitroblue tetrazolium,NBT)阳性细胞百分率和细胞培养上清液中甲胎蛋白(alpha-fetoprotein,AFP)含量,分析CL联合5-FU对SMMC-7721细胞分化的影响;采用丫啶橙(acridineorange,AO)/溴化乙锭(ethidium bromide,EB)荧光染色法和FCM法检测CL联合5-FU对肿瘤细胞周期和细胞凋亡的影响。结果:联合作用组的细胞生长曲线较5-FU和对应浓度CL单独作用组下移,且联合作用组的联合作用系数q值为0.85～1.15,表明CL联合5-FU具有抗肿瘤相加作用;与对照组相比较,5-FU、CL和2药联合均能提高NBT阳性细胞比率(P<0.05),且联合用药组的NBT阳性细胞比率比CL单独作用组高(P<0.05);CL组和CL联合5-FU组均能降低细胞AFP含量,且随着CL浓度的增加而下降,但2组间差异无统计学意义(P>0.05);CL单独作用组和联合作用组间的细胞凋亡率无明显差异(P>0.05);与阴性对照组比较,5-FU组能增加异倍体细胞比率(P<0.05),CL组和CL联合5-FU组均能增加G0/G1期细胞比率(P<0.05)、减少S期细胞比率(P<0.05),联合用药组还能降低G2/M期细胞比率(P<0.05)。结论:CL与5-FU具有协同抗SMMC-7721细胞的作用,其机制可能与2者协同诱导细胞分化和抑制细胞增殖有关。
Objective：To investigate the synergistic antitumor effect of 5-fluorouracil（5-FU）combined with extract from Rosa roxburghii Tratt （CL） on human hepatocellular carcinoma cell line SMMC-7721. Methods：The effect of 5-FU combined with CL on the proliferation of SMMC-7721 cells was measured by MTT reduction test and analyzed by cell growth curves. The effect of 5-FU and CL on the differentiation of SMMC-7721 cells was evaluated by counting the number of nitroblue tetrazolium（NBT）-positive cells and determining the alpha-fetoprotein （AFP） level in cultured supernatant. The effect of 5-FU and CL on cell cycle and apoptosis was determined by acridine orange （AO）/ethidium bromide （EB） staining and flow cytometry （FCM）.Results：The growth curves of SMMC-7721 cells were shifted towards the bottom after combined treatment,and the q values were between 0.85-1.15 in 96 h,indicating there was an additive effect between 5-FU and CL. The ratios of NBT-positive cells were elevated after being treated with either 5-FU or CL alone and in combination compared with control group （P〈0.05）,and the percentage of NBT-positive cells was higher in combined therapy group than single CL therapy group （P〈0.05）. CL or CL combined with 5-Fu decreased the extracellular AFP level. The AFP concentration in cultured medium was decreased with increased CL dosage,but there was no obvious difference between combined treatment group and single CL treatment group （P〉0.05）. The apoptotic ratio was not different between CL and CL plus 5-FU treatment group （P〉0.05）. Compared with negative control group,the ratio of heteroploid was raised after being treated with 5-FU （P〈0.05）. The ratio of SMMC-7721 cells in G0/G1 phase was increased （P〈0.05） and that in S phase was decreased （P〈0.05） after treatment with CL or CL plus 5-FU. The percentage of SMMC-7721 cells in G2/M phase was decreased after treatment with CL plus 5-FU （P〈0.05）. Conclusion：CL and 5-FU have synergistic anticancer effect on SMMC-7721 cells,and the machnisms are related with their synergistic effects in inducing cell differentiation and inhibiting cell proliferation.
出处
《肿瘤》
CAS
CSCD
北大核心
2009年第10期955-960,共6页
Tumor
基金
贵州省科学技术基金资助项目(编号:黔科字3050)
广西教育厅科研资助项目(编号:200810LX450)
