摘要
目的:应用基因芯片技术研究白花蛇舌草豆甾醇(Stigmasterol from Hedyotis diffusa willd.,SHD)抑制人肝癌细胞SMMC-7721生长的靶基因调控。方法:MTT法评价SHD在0、5、10、50、100mg/L浓度下,于24、48、72h对人肝癌细胞SMMC-7721的抑制率变化。分别提取人正常肝细胞、人肝癌SMMC-7721细胞和SHD作用后的SMMC-7721细胞的总RNA,逆转录合成单链、双链cDNA后,体外转录合成生物素标记的cRNA与人HO4基因表达谱芯片杂交,扫描杂交芯片图像,利用软件获得SHD抑制人肝癌的靶基因,对其进行生物信息学分析。结果:SHD对SMMC-7721具有体外抑制作用,且呈剂量依赖性和时间依赖性;SHD使癌基因fos、myc、ras、pim-1、met、rel下调至正常水平,使抑癌基因NF-2和磷酸激酶MAP2K6的表达上调至正常水平。结论:SHD对人肝癌细胞SMMC-7721具有显著的体外抑制作用;SHD抑制SMMC-7721细胞的作用由多条靶基因协同,并通过胞内外信号转导途径协调完成。
Objective： To explore the inhibitory effect of Stigrnasterol from Hedyotis diffusa willd. （SHD）on target gene regulation of human hepatoma cell line SMMC-7721 in vitro by using gene chip technology. Methods： The cultured SMMC- 7721 cells were treated with different concentrations of SHD. The total RNA was extracted from normal human hepatic cells, SMMC-7721 cells and SHD-treated SMMC-7721 cells, and synthesized into double strand eDNA templates. Transcription of cRNA probe with biotin labeling was performed, and then the obtained eDNA was hybridized with human HO4 gene profile. Cy3 dye was detected with Scan array 5000, and the image information was converted into numeric data. Clusting analysis was performed with IMAGENE 1V software. Results： The proliferation of SMMC-7721 cells in vitro was inhibited by SHD with dose and time-dependence. SHD down-regulated the expression of oncogenes, such as fos, myc, ras, pim-1, met, rel, to normal level; it up-regulated anti-oncogenes, such as NF-2 and mitogen-activated protein kinase kinase 6, to normal condition. Conclusion： Hepatoma cell line SMMC-7721 can be inhibited by SHD in vitro.The inhibitory effect induced by SHD relates with various oncogenes and anti-oncogenes.
出处
《现代生物医学进展》
CAS
2007年第8期1181-1183,共3页
Progress in Modern Biomedicine
