新疆阿魏菇粗提物抗肿瘤效应研究

研究新疆阿魏菇粗(水、醇)提物中可能的抗肿瘤有效成分,探讨粗提物抗肿瘤作用的分子生物学机制。方法: 采用天然植物化学分离、分析及鉴定技术,对阿魏菇醇提物进行了有效成分的初步分离、鉴定;运用形态学观察、流式细胞技术及RT-PCR技术,从诱导肿瘤细胞凋亡角度探讨粗提物抗肿瘤机制。结果: 1. 分离所得组分1号、2号及3号结晶可能为同一种物质─山梨醇,4号组分为一氨基酸混合物;2. 分离组分的体外抗肿瘤作用明显低于粗提物;3. 荧光染色实验结果显示,不同剂量的阿魏菇粗提物分别作用12及24 h后,四种类型肿瘤细胞的凋亡细胞百分比均明显高于阴性对照组(P<0.05, P<0.01);4. 流式细胞术及RT-PCR实验结果,(1)阿魏菇醇提物及水提物作用后,Q3、Hela细胞P53蛋白表达水平明显上调,尤以Q3细胞醇提物作用24h最为明显;(2)以-actin的表达及表达量作为内对照,四种类型肿瘤细胞均出现P53蛋白表达水平的提高,p53基因、fas基因mRNA表达水平亦上调(P<0.05, P<0.01)。结论: 阿魏菇粗提物可能通过诱导促进肿瘤细胞凋亡基因(p53基因、fas基因)的转录及表达而发挥抗肿瘤作用,至于p53、fas基因通过何种途径促进肿瘤细胞凋亡,尚待进一步探讨。

STUDY ON THE ANTITUMOR EFFECT OF CRUDE EXTRACT FROM PLEUROTES SAPIDUS

Objective: The antitumor mechanism of Pleurotes sapidus on 4 kinds of tumor cell lines was studied. The active components of its ethanol extracts were studied. Methods:Using techniques of cell culture in vitro, phytochemical methods, FACS and RT-PCR, expressions of p53 and fas gene of four tumor cell lines treated with Pleurotas sapidu were analysed. Results:The apoptosis rates were remarkably increased after treated by ethanol extracts (0.1g/ml) and water extracts (0.1g/ml) for 12 h and 24 h respectively. The expression of P53 protein in the groups of ethanol and water extracts were significantly increased, and the gene expressions of p53 and fas were aslo highly up-regulated. Conclusion:The antitumor effect of Pleurotas sapidus on tumor cell lines in vitro is evident, and its mechanisms are probably associated with the regulation of apoptosis by inducing the gene expressions related to apoptosis.