摘要
目的:观察曲古抑菌素A(trichostatin A,TSA)对甲状腺鳞癌SW579细胞株生长增殖和凋亡的影响,并探讨其作用机制。方法:四甲基偶氮唑蓝(MTT)法检测培养细胞的增殖情况;吖啶橙染色后荧光显微镜下观察细胞凋亡情况;流式细胞术检测细胞凋亡率;RT-PCR方法检测细胞凋亡相关基因caspase-3mRNA表达。结果:TSA明显抑制SW579细胞生长,且呈剂量依赖性。吖啶橙染色后荧光显微镜下观察发现,经TSA处理的各组细胞均可见染色质浓集和边集,核膜出芽及凋亡小体等现象。流式细胞仪分析细胞凋亡率随TSA浓度的升高而升高(P<0.05)。RT-PCR方法检测发现细胞凋亡相关基因caspase-3mRNA表达水平上调。结论:不同浓度的TSA可抑制甲状腺鳞癌SW579细胞增殖,促进细胞凋亡,且呈剂量依赖性;其诱导细胞凋亡的作用机制可能与上调caspase-3基因表达,激活caspase途径有关。
Objective：To study the effects of TSA on proliferation and apoptosis of thyroid squamous cell carcinoma SW579 and its mechanism in vitro. Methods： After 48 hours SW579 cells treated with different dose of TSA （final concentration of 0nmol/L, 25nmol/L, 50nmol/L, 100nmol/L, 200nmol/L, 400nmol/L）, the proliferation activity of cell line in vitro were determined using methyl thiazolyl tetrazolium（MTr） assay. Cell apoptosis was observed by fluorescence microscope stained with acridine orange . Flow cytometry was adopted to examine apoptotic rate. The expression level of caspase - 3 mRNA was determined by RT - PCR. Results ： The proliferation assay analysis showed TSA suppressed significantly the proliferation of cell lines and the effect was in dose dependent, typical apoptotic morphologic feature was discovered under fluorescence microscope stained with acridine orange：chromatin condensation and apoptotic body formation ; The apoptotic rate was increased with the concentration （ The apoptotic rates in SW579 cells were 2.98% in the control group,49.78% in 400nmol/L dose of TSA group）. RT - PCR showed that the expression of caspase -3 mRNA in cells was increased. Conclusion：TSA can inhibite the proliferation and induce apoptosis in thyroid squamous cell carcinoma cell lines SW579 with a dose - dependent manner. The mechanism of TSA inducing cell apoptosis may be related with the facilitating of transcription of caspase -3 and activiting of casepase - 3 pathway.
出处
《现代肿瘤医学》
CAS
2010年第1期32-35,共4页
Journal of Modern Oncology
基金
辽宁省教育厅资助项目(编号:05L132)
