Cui Feng-Jie, Li Yin, Xu Ying-Ying, Liu Zhi-Qiang, Huang Da-Ming, Zhang Zhi-Cai, Tao Wen-Yi
Institute of Bioengineering, School of Food and Biotechnology, Jiangsu University, Xuefu Road, Zhenjiang, Jiangsu 212013, PR China.
Toxicol In Vitro. 2007 Apr;21(3):417-27. doi: 10.1016/j.tiv.2006.10.004. Epub 2006 Oct 13.
The biological function of GFPPS1b, a novel polysaccharide-peptide isolated from cultured mycelia of Grifola frondosa GF9801, was well investigated. GFPS1b has anti-tumor activity and can significantly inhibit the proliferation of SGC-7901 cells, whereas slightly influences the growth of human normal liver cell line L-02. When treated with GFPS1b, SGC-7901 cells showed typical apoptotic morphological features such as the loss of villus and appearance of apoptotic bodies on the cell surface, volume reduction, and chromatin condensation, by scanning electron microscopy (SEM) and fluorescent microscopy (Hoechst 33342). The results of flow cytometry analysis and annexin V-PI assay showed that the SGC-7901 cell cycle was arrested in the G(2)/M phase, the subdiploid peak of DNA characteristic of apoptotic was also observed, and the apoptosis ratio was about 15.08%. DNA isolated from SGC-7901 cells cultured with GFPS1b showed a typical DNA 'ladders' of apoptosis in agarose gel electrophoresis. Further investigation results showed that the apoptotic machinery of SGC-7901 induced by GFPS1b was associated with drop in mitochondrial trans-membrane potential, upregulation of Bax, downregulation of Bcl-2, and activation of caspase-3. Our finding suggests that GFPS1b could suppress SGC-7901 cell growth and reduce cell survival via arresting cell cycle and inducing apoptosis of tumor cells.
对从灰树花GF9801培养菌丝体中分离得到的新型多糖肽GFPPS1b的生物学功能进行了深入研究。GFPPS1b具有抗肿瘤活性,能够显著抑制SGC - 7901细胞的增殖,而对人正常肝细胞系L - 02的生长影响较小。用GFPPS1b处理后,通过扫描电子显微镜(SEM)和荧光显微镜(Hoechst 33342)观察发现,SGC - 7901细胞呈现出典型的凋亡形态特征,如绒毛消失、细胞表面出现凋亡小体、体积缩小以及染色质凝聚。流式细胞术分析和膜联蛋白V - PI检测结果表明,SGC - 7901细胞周期停滞在G(2)/M期,还观察到了凋亡特征性的DNA亚二倍体峰,凋亡率约为15.08%。在琼脂糖凝胶电泳中,从用GFPPS1b培养的SGC - 7901细胞中分离得到的DNA呈现出典型的凋亡DNA“梯状条带”。进一步的研究结果表明,GFPPS1b诱导SGC - 7901细胞凋亡的机制与线粒体跨膜电位下降、Bax上调、Bcl - 2下调以及caspase - 3激活有关。我们的研究结果表明,GFPPS1b可通过阻滞细胞周期和诱导肿瘤细胞凋亡来抑制SGC - 7901细胞生长并降低细胞存活率。