摘要
目的研究姜黄挥发油（turmericvolatileoil，TVO）对人表皮黑素瘤WM35细胞增殖及凋亡的影响，并探讨其作用机制。方法不同浓度（5～160mg／L）TVO体外作用WM35细胞。噻唑蓝（MTT）法测增殖抑制率；倒置显微镜观察细胞形态变化，碘化丙啶（PI）染色，观察荧光数目；DNA片段化检测细胞凋亡；比色法测Caspase-3酶活性；实时荧光定量RT—PCR测Bax及Bcl-2基因mRNA表达水平；流式细胞仪测细胞凋亡及周期。结果TVO对WM35细胞有抑制增殖的作用，且呈时间一剂量依赖性（P〈O．05）。TVO诱导WM35细胞凋亡，呈剂量依赖性（P〈0．05），Caspase-3酶活性随药物浓度增加而增强（P〈0．05），Bax／Bcl-2表达水平比值随药物浓度增加而升高（P〈0．05）；细胞周期阻滞在s期。结论TVO对WM35细胞有抑制增殖及促进凋亡的作用，其机制是使凋亡相关基因Bax表达水平上调、Bcl-2表达水平下调，激活细胞凋亡途径关键酶Caspase-3，将细胞周期阻滞在s期，进而抑制肿瘤细胞分化与增殖。
Objective To study the effect of turmeric volatile oi1（TVO） on proliferation and apoptosis in human mela- noma cell line WM35 cells,and the possible mechanisms. Methods WM35 cells were incubated with differ- ent concentrations of TVO （5 x 160mg/L）. Cell proliferation was measured by MTF assay. Inverted micro- scope was used to assess the cell morphology and the number of PI staining positive cells. Apoptosis was de- tected by DNA fragmentation. Caspase-3 activities were measured using a colorimetric method. The expression levels of apoptosis-associated mRNA （ Bcl-2 and Bax） were determined using RT-PCR. The apoptosis and cell cycle were analyzed by flow cytometry. Results TVO dose-dependently inhibited WM35 cell prolifera- tion（ P 〈 0. 05 ）, and induced apoptosis（ P 〈 0. 05 ）. Likewise ,TVO also increased Caspase-3 activity and the expression levels of Bax/Bcl-2 mRNA in a dose-dependent manner （ P 〈 0. 05 ）. Cells were arrested in S phase after TVO treatment. Conclusion TVO-induced inhibition of WM35 cell proliferation and apoptosis are likely via upregulating Bax and down-regulating Bcl-2 expression. Retention of cells in S phase and acti- vation of Caspase-3 could contribute to TVO-induced inhibition of tumor cell proliferation and differentiation.
出处
《中国皮肤性病学杂志》
CAS
CSCD
北大核心
2015年第12期1227-1232,共6页
The Chinese Journal of Dermatovenereology
基金
贵州省中药现代化专项项目[黔科合中药字(2012)5018号]
贵阳市科技局科技创新平台项目[筑科合同(2012303)号]
