摘要
目的研究TOFA及柔红霉素（DNR）联合应用对肠癌细胞株HCT-8的作用，探讨其联合应用于临床的可能性。方法将1×10^4个HCT-8细胞种植于96孔板，采用MTT比色法测定不同浓度TOFA或／和DNR对同步化处理后的HCT-8细胞的生长抑制效果；DNA降解分析法（DNA Fragmemation）检测TOFA与DNR联合使用对HCT-8细胞DNA的损伤。Westernh10t检测TOFA与DNR联合使用对凋亡蛋白p53以及PARP的水解作用。结果单用TOFA或DNR均可呈浓度依赖性抑制肠癌细胞HCT-8的生长，其IC50值分别为7．5μg／mL和0．18μmoL／L；当用20μg/mLTOFA或0．6μmol／LDNR处理时，活性细胞只有正常对照组（无药物处理组）的26．2％±5．1％或22．3％±4．5％。当用20μg/mLTOFA和0．6μmol/LDNR两药联合应用时，活性细胞只有正常值的10．4％±3．0％。同时联合应用可显著增加肿瘤细胞DNA的降解；并显著诱导凋亡蛋白p53表达，促进PARP的水解。结论TOFA与DNR联合应用可协同抑制肠癌细胞株HCT-8细胞的生长，其效应与药物诱导细胞凋亡有关。
Objective To investigate the effect of combination application of TOFA and Daunorubsin （DNR） on cell growth of colon cancer cell line HCT-8. Methods 1 x 104 cells of HCT-8 each well were spread in 96-well plate and cultured overnight. The cells were treated with serum-free RPMI-1640 medium for 24 h for cell synchronization, and then with different concentrations of TOFA or DNR for 72 h. Finally, viable cells were detected by M3T assay. After HCT-8 cells were treated with 20 g/mL TOFA and 0.6μmol/L DNR for 24 h, DNA was extracted from HCT-8 cells and subjected to DNA fragmentation, and protein was extracted from HCT-8 and subjected to western blot for detecting p53 and PARP. Results TOFA or DNR inhibited the growth of HCT-8 in a dose-dependent manner. TOFA and DNR are cytotoxic to HCT-8 cells with an ICs0 at approximately 7.5μg/mL or 0.18μmol/L,respectively. When TOFA and DNR with the concentrations of 20 pLg/mL or 0.6 μmol/L were associately applied, the cell viability was only 10.4% ± 3.0%, contrast to control without drug treatment. The combination application of the two drugs significantly increased DNA fragmentation, and also induced the expression of p53,and the cleavage of PARP. Conclusion The combination applicaiton of TOFA and DNR inhibits the growth of colon cancer cell line HCT-8 by inducing cell apoptosis.
出处
《中南医学科学杂志》
CAS
2011年第5期514-517,521,共5页
Medical Science Journal of Central South China
基金
国家自然科学基金(30971170)
湖南省自然科学基金项目(06JJ5055)
