南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (01): 75-79.

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一种新的蝮蛇蛇毒磷脂酶A2同源物的分离纯化及其对Hep3B细胞基因谱的影响

马安德;吴少瑜;张嘉杰;李志琴;徐伟;文晓芸;余乐;吴曙光;   

  1. 南方医科大学分析测试中心； 南方医科大学药学院； 南方医科大学药学院 广东 广州 510515； 广东 广州 510515；
  • 出版日期:2006-01-20 发布日期:2006-01-20
  • 基金资助:
    广东省重点课题(2005B30101006)

Purification of a new phospholipase A2 homologue from Agkistrodon blomhoffii siniticus and its effects on gene expression profile of Hep3B cells

MA An-de, WU Shao-yu, ZHANG Jia-jie, LI Zhi-qin, XU Wei, WEN Xiao-yun, YU Le, WU Shu-guang Instrumental Analysis and Research Center, School of Pharmacological Sciences, Southern Medical University, Guangzhou 510515, China   

  1. 南方医科大学分析测试中心； 南方医科大学药学院； 南方医科大学药学院 广东 广州 510515； 广东 广州 510515；
  • Online:2006-01-20 Published:2006-01-20

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摘要： 目的从中国蝮蛇蛇毒中分离纯化出一种新的磷脂酶A2(PLA2)同源物,并研究其对Hep3B细胞基因谱的影响。方法用C18反相高效液相层析从蛇毒粗毒中分离纯化出PLA2同源物,并检测其纯度。用电喷雾质谱仪测定PLA2 同源物分子量。体外培养Hep3B细胞,以139μg／ml PLA2同源物处理12 h,应用基因芯片检测基因表达的改变。结果从蛇毒粗毒中分离纯化出一种新的PL,A2同源物,纯度为97．2％,相对分子质量为13 900。139μg／ml PLA2同源物处理 Hep3B细胞12 h后,19个基因表达下调,20个基因表达上调。表达改变的基因按功能分主要为以下6类:细胞周期控制和DAN损伤修复类、细胞调亡和衰老类信号转导分子和转录因子、细胞黏附类、血管生成类以及肿瘤细胞入侵和转移类。结论PLA2同源物作用于Hep3B细胞后,可影响细胞多种基因的表达,表达改变的基因主要参与肿瘤细胞的生长和转移。本研究为进一步深入研究PLA2对肿瘤细胞的作用机制提供线索和依据。 更多还原

Abstract: Objective To isolate and purify a new phospholipase A2 (PLA2) homologue from Agkistrodon blomhoffii siniticus and investigate its effects on the gene expression profile of Hep3B cells. Methods The PLA2 homologue was isolated and purified by reverse-phase high-performance liquid chromatography (HPLC) and its purity was determined also by HPLC. The relative molecular mass of the homologue was measured by electrospray ionization mass spectrum. The gene expression profile of Hep3B cells was detected with gene chip after exposure of the cells to 139 μg/ml PLA2 homologue for 12 h. Results The purity of the PLA2 homologue was 97.2%, whose relative molecular mass was 13 900. After exposure of Hep3B cells to 139 μg/ml PLA2 homologue for 12 h, 19 genes were down-regulated and 20 up-regulated in the cells. The genes showing altered expressions in response to the exposure were mainly involved in cell cycle control and DNA damage repair, cell apoptosis and senescence, production of signal transduction molecules and transcription factors, cell adhesion, angiogenesis, and tumor invasion and metastasis. Conclusions The PLA2 homologue induces alterations in the expression of a wide variety of genes involved in the growth and metastasis of tumor cells. The results of this study provide clues for further study of the possible mechanism for the action of PLA2 homologue on Hep3B cells.

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