This work was supported by the National Natural Science Foundation of China (No. 30371753), the Research Project of Hebei Education Department (No. 2004110) and Doctor Foundation of Hebei Normal University (No. L2005B25).

^*Corresponding author. Tel: +86-311-6268655; Fax: +86-311-86268313; E-mail: ligao0823@163.com; gaoli0823@yahoo.com.cn

Effect of the venom of the spider Macrothele raveni on the expression of p21 gene in HepG2 cells

GAO Li^1,*, SHEN Jin-Bao², SUN Jie³, SHAN Bao-En⁴

¹College of Life Science, Hebei Normal University, Shijiazhuang 050016, China; ²Huihua College, Hebei Normal University, Shijiazhuang 050011, China; ³Science and Technology company, Hebei Medical Unversity, Shijiazhuang, 050017, China; ⁴Research Centre, The Fourth Hospital of Hebei Medical University, Shijiazhuang, 050011, China

Abstract: This paper focuses on the effect of the venom of the spider Macrothele raveni on the proliferation of human hepatocelluar carcinoma cell line HepG2 and the related molecular mechanism. MTT test showed that the proliferation of HepG2 cells in vitro was inhibited by the spider venom (P<0.05) in a concentration-dependent manner. By using flow cytometry it was found that the spider venom caused selective G₂/M cell cycle arrest in HepG2 cells. RT-PCR and Western blot indicated the expressions of p21 mRNA and protein in HepG2 cells were obviously up-regulated by the spider venom. The venom of the spider Macrothele raveni inhibited the proliferation of HepG2 cells. These results suggest that the possible mechanism of venom is to activate the expression of p21 gene and protein and to cause selective cell cycle arrest in G₂/M, leading to HepG2 cell apoptosis.

Key words: the venom of the spider Macrothele raveni; HepG2 cells; cell cycle; p21 gene; p21 protein

雷氏大疣蛛毒液对人肝癌HepG2细胞p21基因表达的影响^*

高莉^1,*，沈金宝² ，孙 捷³，单保恩⁴

¹河北师范大学生命科学学院，石家庄 050016；²河北师范大学汇华学院，石家庄 050091；³河北医科大学科技总公司，石家庄 050017；⁴河北医科大学第四医院科研中心，石家庄050011

摘 要：本文研究了雷氏大疣蛛毒液对人肝癌细胞株HepG2增殖抑制作用及其分子机制。采用MTT法观察到，雷氏大疣蛛毒液剂量依赖抑制HepG2 细胞增殖；流式细胞仪检测发现，经过雷氏大疣蛛毒液作用的HepG2细胞周期发生选择性的明显改变；RT-PCR方法检测到p21基因表达增强；Western blot发现，P21蛋白表达增加。结果提示，雷氏大疣蛛毒液抑制人肝癌细胞HepG2增殖的可能机制之一是使p21基因和蛋白表达增加，G₂/M 细胞周期被阻滞，从而诱导细胞的凋亡。

关键词： 雷氏大疣蛛毒液；HepG2细胞；细胞周期；p21基因；p21蛋白

中图分类号：R33