摘要
背景与目的:探讨莱菔硫烷对人膀胱癌细胞(T24)细胞周期的影响及作用机制。材料与方法:采用噻唑蓝(MTT)法研究不同浓度的莱菔硫烷对T24细胞生长的抑制作用并测定其半数抑制浓度(IC50);采用流式细胞仪检测莱菔硫烷对T24细胞周期的影响;采用westernblot研究莱菔硫烷对T24细胞中细胞周期蛋白依赖激酶抑制剂P16和P27的表达情况。结果:在较低剂量范围内(≤40μmol/L),随着作用剂量的增加,莱菔硫烷对T24细胞的生长的抑制作用也明显增强。10、20、40μmol/L莱菔硫烷的抑制率分别为(12.5±3.95)%,(25.0±2.50)%、(50.0±5.33)%;在较高剂量(60μmol/L^160μmol/L)时,这种抑制作用不再呈剂量依赖性;莱菔硫烷作用72h后的IC50值为(51.12±7.10)μmol/L;莱菔硫烷能使T24细胞周期阻滞于G0/G1期,20μmol/L莱菔硫烷作用48h后,在G0/G1峰之前出现凋亡峰;20μmol/L莱菔硫烷作用于T24细胞8、12、24h后能明显诱导P27蛋白的表达,作用早期(8h)时能诱导P16蛋白的表达。结论:莱菔硫烷能抑制T24细胞生长并使该细胞周期阻断在G0/G1期,其作用机制主要是通过诱导P27蛋白及早期诱导P16蛋白来实现的。
BACKGROUND ＆AIM： To study the effects and mechanisms of sulforaphane on T24 cell cycle. MATERIALS AND METHODS： The inhibitory effects of sulforaphane on the growth of T24 cells were investigated and its value of IC50 after 72 h incubation were measured by 3-（4,5-dimethyl-thiazolyl-2）-2,5-diphenyl tetrazolium bromide（MTT） method. The effect of sulforaphane on the T24 cell cycle was determined by flow cytometry; and expressions of P27 and P16 induced by sulfomphane were studied by westem blotting analysis. RESULTS： Sulfomphane in low concentrations （≤40 μmol/L）could significantly suppress the growth of T24 cells in a dose-dependent way.The inhibitory rates for 10 μmol/L,20 μmol/L and 40 ttmol/L were （12.5±3.95）%, （25.0±2.50）% and （50.0±5.33）%,respectively. High concentrations of sulfomphane（60 μmol/L- 160 μmol/L）, the inhibitory effects showed no obvious dose-dependency. The IC50 value of sulforaphane treatment for 72 h was （51.12±7.10）ttmol/L. Sulforaphane could block T24 cell cycle at the G0/G1 phase as showed by flow cytometry. Moreover, the treatment of 20 μmol/L sulfomphane for 48 h caused the appearance of pre-G1 before G0/G1 phase. Treatments with 20 μmol/L sulfomphane for 8, 12, 24 h significantly induced the expression of P27 protein in a time-dependent manner. At early stage of sulforaphane treatment（8 h） , the expression of P16 protein was also induced. CONCLUSION： Sulfomphane could inhibit the growth of T24 cells and block cell cycle at G0/G1 phase. The induction of P27 protein by sulfomphane mainly involved molecular mechanisms, may be including the early induction of P16 protein.
出处
《癌变·畸变·突变》
CAS
CSCD
2006年第6期443-446,共4页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
黑龙江省卫生厅资助项目(No.2005-08)
