| 摘要: |
| 目的:观察来源于海芒果种子的强心苷化合物β-D-glucosyl-(1-4)-α-L-thevetosides of 17β-digitoxigenin(GHSC-73)对人肝癌细胞株HepG2细胞增殖和细胞周期的影响，并初步探讨其作用机制。方法:采用MTT法检测不同浓度GHSC-73对人肝癌细胞株HepG2增殖的影响；采用流式细胞仪Propidium Iodide(PI)单标法检测GHSC-73对HepG2细胞周期进程的影响；Real-time RT-PCR检测S期相关基因在GHSC-73处理前后表达的变化。结果:GHSC-73可抑制HepG2细胞增殖，其作用具有时间和浓度依赖性,作用HepG2细胞24、48、72 h后,IC50分别为(5.18±0.21)、(0.37±0.08)、(1.66±0.16) μmol/L。与对照组相比，随着作用时间的延长，S期细胞百分比逐渐增多，而G0/G1期细胞百分比逐渐减少(P<0.05),G2/M期细胞百分比基本保持不变，表明GHSC-73阻滞HepG2细胞于S期。Real-time RT-PCR检测S期相关基因的结果显示：GHSC-73诱导后HepG2细胞GADD153、cyclin D1、p21基因表达上调，cyclin A2、 DHFR、TYMS基因表达下调。结论:GHSC-73可通过S期阻滞来抑制HepG2细胞增殖，其阻滞作用可能与GADD153、cyclin D1、 p21、 cyclin A2、DHFR和TYMS基因表达变化有关。 |
| 关键词: 肝肿瘤 强心苷类 GHSC-73 细胞增殖 细胞周期 |
| DOI：10.3724/SP.J.1008.2009.01110 |
| 投稿时间：2009-03-23修订日期：2009-09-18 |
| 基金项目:国家高技术研究发展计划(“863”计划，2006AAD9Z447). |
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| Effects of GHSC-73 on proliferation and cell cycle of human hepatocellular carcinoma HepG2 cells |
| FENG Bo1,GUO Yue-wei2,HUANG Cai-guo1*,LI Liang2,JIAO Bing-hua1* |
| (1.Department of Biochemistry and Molecular Biology,College of Basic Medical Sciences,Second Military Medical University,Shanghai 200433,China;2.State Key Laboratory of Drug Research,Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai 201203) |
| Abstract: |
| Objective:To investigate the effects of β-D-glucosyl-(1-4)-α-L-thevetosides of 17β-digitoxigenin (GHSC-73),an isolate from the seeds of Cerbera manghas L.,on cell growth and cell cycle regulation of human hepatocellular carcinoma cell line HepG2,and to discuss the related mechanism.Methods: HepG2 cells were treated with different concentrations (0-80 μmol/L) of GHSC-73.Cell viability was determined using MTT assay at 24,48,and 72 h after treatment.Cell cycle distribution was assessed by flow cytometry after propidium iodide (PI) staining.Expression of the S phase associated genes GADD153,cyclin D1,cyclin A2,DHFR,TYMS,and p21 was determined by real-time RT-PCR before and after GHSC-73 treatment.Results: GHSC-73 inhibited the cell proliferation of HepG2 cells in a dose- and time-dependent manner.The values of IC50 were (5.18±0.21),(0.37±0.08),and (1.66±0.16) μmol/L at 24,48,and 72 h,respectively.Compared with control group,the proportion of cells in S phase increased in the treatment group with the prolongation of treatment,the cells in G0/G1 phase gradually decreased (P<0.05),and cells in G2/M phase remained unchanged,indicating that GHSC-73 blocked HepG2 cell in S phase.Real-time RT-PCR showed that cyclin A2,DHFR, and TYMS were down-regulated,and p21,GADD153, and cyclin D1 were up-regulated,which might be associated with the GHSC-73-induced S phase arrest in HepG2 cells.Conclusion: GHSC-73 can inhibit growth of HepG2 cells by inducing S phase arrest; down-regulation of cyclin A2,DHFR, and TYMS genes and up-regulation of p21,GADD153,and cyclin D1 genes might participate in the induction of arrest. |
| Key words: liver neoplasms cardiac glycosides GHSC-73 cell proliferation cell cycle |
