MTT法指导分离壁虎抗肿瘤活性成分的实验研究

目的 采用四氮唑盐（MTT）法指导壁虎抗肿瘤活性成分的分离,寻找壁虎抗肿瘤活性单体化合物,并初步观察其抗肿瘤作用.方法 常规培养人肝癌Bel-7402细胞、人肺癌Bel-95C细胞,在倒置显微镜下观察细胞形态及数量,采用MTT法检测多批次壁虎活性成分提取物对体外培养的肿瘤细胞株的生长抑制作用,计算肿瘤抑制率.结果 经过11批次重复药筛实验,逐步分梯次指导分离纯化,得到壁虎抗肿瘤活性单体化合物＂守宫咪唑衍生物＂.该化合物可明显抑制人肝癌Bel-7402细胞的生长,用药后前3d的抑瘤率分别为71.24%,91.37%,94.45%,在显微镜下发现肿瘤细胞变圆、体积变小.结论 用MTT法结合倒置显微镜观察细胞状态,指导中药壁虎抗肿瘤活性成分的提取分离,得到抗肿瘤作用明显的壁虎单体化合物.

Experimental Study of Separating Gekko Swinhonis Active Constitutent Using MTT Method in Vitro

Objective To separate Gekko Swinhonis anti-tumor active constitutent in vitro by MTT assay. To search Gekko Swinhonis anti-tumor active monomer compounds, as well as to observe the effect of anti-tumor preliminarily. Methods Human hepatoma Bel-7402 cells and human lung cancer cells Bel-95C were cultured conventionally. Then cell morphology and the number of cells were observed by inverted microscope. The growth inhibition effects of multiple Gekko Swinhonis anti-tumor active constitutent extractions on tmnor cells cuhured in vitro were detected by MTT assay. And tumor control rates were calculated. Results After 11 batches of repeated medicinal sieve tracer, separation and purification were directed progressively. Gekko Swinhonis anti-tumor activity monomer compound named ＂Gekko Swinho- nis Imidazole derivatives＂ was obtained uhimately. The compound couhl inhibit human hepatocellular carcinoma Bel- 7402 cells obviously. The tumor inhibition rates of the first 3 days were 71.24% ,91.37% and 94.45% respectively. Tumor cells became round and smaller under microscope. Conclusion Cell state was observed well using MTT method combined with inverted microscope. Then the separation and purification of Gecko anti-tumor active compound were traced. Gekko Swinhonis anti-tumor active monomer compound with obvious anti-tumor effect was obtained uhimately.