摘要
【目的】探讨PPAR-γ选择性抑制剂T0070907影响鼻咽癌(NPC)细胞体外生长的作用机制。【方法】通过RT-PCR和Westernblotting检测PPAR-γ受体在5株NPC细胞株(CNE1、CNE2、HONE1、SUNE1和5-8F)中mRNA和蛋白水平的表达;MTT法检测T0070907对NPC细胞生长情况的影响;流式细胞术和Westernblotting检测T0070907对NPC细胞的细胞周期和细胞周期相关蛋白表达的影响。【结果】PPAR-γ在5株NPC细胞中均有表达;T0070907对NPC细胞株有明显生长抑制作用,并呈浓度、时间依赖性;T0070907作用于NPC细胞CNE1和CNE2后,G2/M期细胞比例则逐渐增加,且随着处理浓度的增加而增加,各组间差异有统计学意义(P<0.05)。此外,随着T0070907处理浓度的增加,NPC细胞的细胞周期相关蛋白CyclinA、CyclinB1、Cdc2、Cdk2蛋白的表达逐渐下降,无活性的pCdc2蛋白表达则逐渐增强。【结论】PPAR-γ抑制剂T0070907能够抑制NPC细胞PPAR-γ的蛋白表达,并通过对细胞周期相关蛋白CyclinA、CyclinB1、Cdc2、Cdk2、pCdc2等蛋白的调控导致细胞周期G2/M期阻滞,从而抑制NPC细胞的生长。
【Objective】PPAR-γ（peroxisome proliferator-activated receptor-γ）antagonists have been documented to induce cancer cell proliferation inhibition and apoptosis.In the present study PPAR-γ＇s effect on cell growth of NPC cell lines and the possible mechanism was investigated via its selective inhibitor T0070907.【Methods】PPAR-γmRNA and protein expression in five human NPC cell lines（CNE1,CNE2,HONE1,SUNE1,and 5-8F）was detected with RT-PCR and Western blotting analysis.NPC cells growth inhibition by T0070907 was measured by MTT assay,and cell cycle arrest of NPC cells treated with T0070907 was assayed by fluorescence-activated cell sorter（FACS）analysis.Cell cycle-associated protein expression was detected by Western blotting.【Results】PPAR-γwas expressed both at mRNA and protein levels in five NPC cell lines.NPC cells＇proliferation was significantly inhibited by T0070907,and the growth inhibition was dose-and time-dependent.CNE1 and CNE2 cells treated for 48 h with T0070907 markedly accumulated dose-dependently in the phase of G2/M,while they differed significantly（P0.05）in the phase as well.T0070907 suppressed expression of cell cycle-related proteins including Cyclin A,Cyclin B1,Cdc2,and Cdk2 but increased pCdc2 expression.【Conclusion】PPAR-γinhibitor T0070907 could induce proliferation inhibition of NPC cells to the phase of G2/M,and this cell cycle arrest might result probably from regulation of Cyclin A,Cyclin B1,Cdc2,Cdk2,and pCdc2.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2010年第3期343-349,共7页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省自然科学基金(031726)
