松针不同提取部位体外抗肿瘤作用的实验研究01.doc

松针不同提取部位体外抗肿瘤作用的实验研究 周微a，郑晓珂b，王小兰a，冯卫生a （河南中医学院，a．药学院；b．基础医学院，郑州 450008） 摘 要：目的 探讨松针不同提取部位的体外抗肿瘤作用。方法 采用系统溶剂萃取法将松针醇提物进行部位分离，得到石油醚、乙酸乙酯、正丁醇及水提取部位，通过四甲基偶氮唑盐比色法（MTT法）观察各提取部位对人肝癌细胞株SMMC-7721、HepG2MCF-7，人前列腺癌细胞株PC-3细胞增殖的抑制作用。采用流式细胞术分析细胞周期分布，初探松针抗肿瘤作用机理。结果 石油醚部位对肿瘤细胞的抑制作用较强，在一定剂量范围内呈现出时间和剂量依赖性；乙酸乙酯部位的作用次之；正丁醇和水部位的抑制作用不明显。SMMC-7721细胞经0.6 mg·ml-1、0.8 mg·ml-1石油醚提取部位处理48h后的凋亡百分率分别为18.7%、21.4%，它对该细胞增殖的抑制是将细胞周期阻滞在G2-M期。结论 松针提取物在体外具有抗肿瘤作用，石油醚部位呈现出较强的抑制作用，其体外抑制SMMC-7721细胞增殖的机制可能与松针提取物的细胞毒作用、诱导细胞凋亡和抑制细胞分裂有关。 关键词：松针；肿瘤细胞株；MTT法；流式细胞术 Experimental Study on Antitumor Effect of Different Fractions of Pine Needles in vitro ZHOU Weia,ZHENG Xiao-keb,WANG Xiao-lana,FENG Wei-shenga (Henan University of Traditional Chinese Medicine,a.School of Pharmacy;b.Basic Medical College,Zhengzhou 450008) Abstract: Objective To study the antitumor effect of different fractions of pine needles in vitro. Methods Pine needles were extracted and separated by the technique of solvent-refining.MTT assay was used to observe the inhibition of extracts on the proliferation of human hepatocellular carcinoma SMMC-7721, HepG2, breast carcinoma MCF-7 and prostatic carcinoma PC-3.Apoptosis rates and cell cycle distribution were detected by flow cytometry analysis.Results Petroleum ether extract from pine needles possesses the role of antitumor effect in cell culture. The inhibitory effect on the proliferation of tumor cells has remarkable time-dependence and dose-dependence. Petroleum ether extract induces SMMC-7721 apoptosis and arrests in G2-M phase . Apoptotic percentage of Petroleum ether extract 0.6mg·ml-1 and 0.8mg·ml-1 on SMMC-7721 is 18.7% and 21.4% in 48 hours. Conclusion Pine needles fractions have the antitumor effect in vitro and the petroleum ether extract possesses the role of antitumor effect.Anticancer mechanism of petroleum ether extract on SMMC-7721 may be direct cytotoxic effect,inducing cell apoptosis and inhibiting cell segmentation. Key words: Pine needles; carcinoma cell lines; MTT; flow cytom