摘要
目的研究manumycin对乳腺癌腹腔转移癌细胞株SK-BR-3的抑癌效应及其诱导凋亡。方法用MTT法检测manumycin对SK-BR-3细胞的抑癌作用。免疫印迹方法检测p38MAPK蛋白表达。用caspase-3活性检测试剂盒定量检测manumycin诱导细胞凋亡的水平及评估特异性的p38MAPK抑制剂SB203580对凋亡的影响。结果经6μmol/L、18μmol/L、54μmol/Lmanumycin处理SK-BR-3细胞24h时,其抑制率分别为(74±39)%、(210±44)%和(647±41)%,呈量效关系。其中后2者的细胞活性与对照组比有显著差异(P<001)。用药24h的IC50为425μmol/L。同时此药物可明显增加caspase-3的活性,且这一效应可部分地被p38抑制剂SB203580阻断。免疫印迹结果显示manumycin促进p38的磷酸化。结论manumycin可通过诱导SK-BR-3细胞凋亡而产生抑癌作用,p38MAPK是manumycin诱导细胞凋亡的通路之一。
AIM: To investigate the anticancer effect of manumycin on abdominal metastatic breast cancer cell line-SK-BR-3 and its relationship with p38 MAPK . METHODS: The test of anticancer effect was performed by the method of MTT, apoptosis induced by manumycin and affected by SB203580, a specific p38 MAPK inhibitor, were examined by caspase-3 activity assay kit, and the protein expression was detected by immunoblotting assay. RESULTS: The inhibition rates at 24 h after treatment with manumycin of 6 μmol/L, 18 μmol/L, 54 μmol/L were (7.4±3.9)%, (21.0±4.4)% and (64.7±4.1)%, respectively and showed dosage-effect relationship. Compared with the control group, the survival rates of the last two treatment groups were decreased significantly (P<0.01). The value of IC 50 24 h after treatment with manumycin was 42.5 μmol/L. Manumycin simultaneously activated caspase-3 protein, which was partly blocked by p38 MAPK inhibitor, SB203580. The results of immunoblotting showed that manumycin increased p38 MAPK protein phosphorylation. CONCLUSION: Manumycin exerts anticancer effect on SK-BR-3 cell line via inducing cell apoptosis, which is partly regulated by p38 MAPK . [
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2004年第12期2311-2315,共5页
Chinese Journal of Pathophysiology
