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目的:通过体外抗肿瘤实验选择鼠妇抗肿瘤活性物质的提取方法和敏感细胞株,为筛选抗肿瘤活性物质做准备。方法:采用水回流提取、水浸渍提取、95%乙醇回流提取、乙醇浸渍提取、仿生酶解提取的方法对鼠妇粉末进行处理,并将其提取物对人肺腺癌细胞(A549)、肝癌细胞(BEL-7402)、乳腺癌细胞(MCF-7)和喉癌细胞(Hep-2)5种癌细胞进行体外抗肿瘤实验,同时采用MTT细胞增殖检测的方法对其抗肿瘤效果进行追踪,确定最佳提取方法和敏感细胞株后,对该方法的提取物用氯仿、正丁醇进一步萃取,再将各萃取部位用敏感细胞株分别进行体外抗肿瘤实验,筛选出最佳抗肿瘤活性部位。结果:与其余4种方法的提取物相比,乙醇回流提取物对A549、BEL-7402、MCF-7、Hep-2 4种癌细胞的抑制作用最好,IC50分别为185.99、119.80、52.94、193.08μg/m L,该提取物的氯仿萃取物对MCF-7的抑制作用明显高于正丁醇萃取物,IC50分别为58.13、190.47μg/m L。结论:乙醇回流提取物对多种癌细胞均有抑制作用,其抗肿瘤活性物质集中在氯仿萃取物中。
OBJECTIVE: To select the extraction method of antitumor active substances and sensitive cell lines in vitro by anti-tumor experiments in mice and prepare for the screening of anti-tumor active substances. Methods: Murine powder was treated with water reflux extraction, water immersion extraction, 95% ethanol reflux extraction, ethanol extraction and biomimetic enzymatic extraction. The extracts were treated with human lung adenocarcinoma cell line A549, The antitumor effects of 5 kinds of cancer cells including BEL-7402, MCF-7 and Hep-2 were tested in vitro and the antitumor effects were tracked by MTT cell proliferation assay. After the optimal extraction method and sensitive cell line were identified, the extract of this method was further extracted with chloroform and n-butanol, and each of the extracted parts was subjected to in vitro anti-tumor experiments with sensitive cell lines to screen out the best anti-tumor active site . Results: Compared with the other four extracts, ethanol extract had the best inhibitory effect on four kinds of cancer cells A549, BEL-7402, MCF-7 and Hep-2 with IC50 of 185.99, 1199.80 and 52.94, 193.08μg / m L, the extract of chloroform extract MCF-7 inhibition was significantly higher than the n-butanol extract, IC50 were 58.13,190.47μg / m L. CONCLUSION: Ethanol reflux extract has inhibitory effect on many kinds of cancer cells, and its anti-tumor active substance is concentrated in chloroform extract.