目的 MTF法是评价细胞增殖的常规方法,但有时有一定的假阳性率,本文从细胞衰老角度进行研究,探讨假阳性率产生的原因和改进方法.方法 使用能引起细胞衰老的抗肿瘤药物博来霉素和博宁霉素,处理人子宫颈癌HeLa细胞和人乳腺癌MCF-7细胞,采用MTT、SRB、台盼蓝染色计数法和克隆形成率等方法检测细胞增殖的抑制作用;衰老相关的β-半乳糖苷酶染色确定衰老细胞.结果 低浓度的博宁霉素处理MCF-7细胞,明显引起细胞衰老.MTT法测得的IC50值最大,SRB和台盼蓝染色计数法测得的IC50值次之,克隆形成率法最为敏感.测得的IC50值最小,达到了nM级.结论 细胞衰老明显影响一些抗肿瘤药物的MTT评价结果,使用克隆形成率法是最为可靠的克服细胞衰老影响的方法.%Objective MTT assay is a conventional method to detect the growth-inhibitory effect. But sometimes there are some false-positive results. This study was to explore the causes of false-positive results from the cellular senescence. Methods HeLa and MCF-7 were detected by MTT, SRB, trypan Blue-stained counting and colony formation assay after exposure to bleomycin and boningmycin which induced cellular senescence in human tumor cells. Cellular senescence was detected with senescence-associated beta-galactosidase staining. Results The IC60 value measured by MTT assay was the largest and the IC50 by clone formation assay was the most sensitive, reaching nanomolar level. Conclusions Cellular sen-escence affects the accuracy of the results of MTT assay. Colony formation assay is the most reliable method to overcome the effects of cellular senescence.
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