摘要
目的研究氯化两面针碱(nitidine chloride,NC)处理后p53通路的分子水平变化,以说明p53途径在NC抑制人肝癌细胞SMMC-7721增殖中的作用。方法应用MTT比色法测定NC对SMMC-7721细胞存活率的影响,细胞克隆集落形成实验测定NC对细胞增殖的抑制作用;实时荧光定量PCR检测不同剂量NC对p53、p21mRNA表达的影响;酶联免疫吸附法测定p53、p21蛋白含量的变化。结果 NC(0.15,0.30,0.60,1.20,2.40,4.80 mg.L-1)浓度依赖性地降低SMMC-7721的存活率,作用48 h时IC50值为(1.03±0.18)mg.L-1;NC可明显抑制细胞集落形成,剂量为0.15,0.30 mg.L-1时,抑制率分别是74.65%、88.48%,剂量为0.60 mg.L-1时,抑制率达到了100%;NC可明显提高癌细胞中p53、p21 mRNA和蛋白的表达,并呈浓度依赖性。结论 p53通路可能在NC诱导的SMMC-7721细胞增殖抑制中起重要作用。
Objective To study the molecular changes of p53 pathway in SMMC-7721 cells treated with nitidine chloride（NC） and to illustrate the role of p53 pathway in NC inhibiting the proliferation of SMMC-7721 cells. Methods Methyl thiazolyl tetrazolium （MTY） colorimetry was applied to evaluate the survival rate of SMMC-7721 cells, and the cell colony formation test was used to detect the anti-proliferation effect of NC on SMMC-7721 ceils. P53 and p21 mRNA levels were quantified by real-time fluorescence quantitative PCR after detecting their protein expression levels by ELISA. Results NC at the concentrations of 0.15, 0.30, 0.60, 1.20, 2.40 and 4.80 mg·L-1 decreased the survival rate of SMMC-7721 cells in a concentration-dependent manner, and the 48 h IC50 value was（1.03± 0.18） mg·L-1. NC obviously inhibited the formation of cell colonies of SMMC-7721 ceils. The colony formulation rate was 74.65 %, 88.48 % and 100 % respectively in the groups of NC at the concentrations of 0.15, 0.30 and 0.60 mgL-1. The protein and mRNA expression levels of p53 SMMC-7721 cells were exposed to NC. Conclusion induced proliferation of SMMC-7721 cells. and p21 in SMMC-7721 cells were obviously up-regulated after P53 pathway probably plays important roles in inhibiting NC-induced proliferation of SMMC-7721 cells.
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2011年第4期385-390,共6页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金(30760302)
广西研究生教育创新计划课题(2009105981007D25)
