南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2013年
10期
1409-1415
,共7页
曲古抑菌素A%帕比司他%肾癌细胞%凋亡%细胞周期%组蛋白去乙酰化酶抑制剂
麯古抑菌素A%帕比司他%腎癌細胞%凋亡%細胞週期%組蛋白去乙酰化酶抑製劑
곡고억균소A%파비사타%신암세포%조망%세포주기%조단백거을선화매억제제
apoptosis%cell cycle%histone deacetylase inhibitor%human renal cell carcinoma%LBH589%trichostatin A
目的研究组蛋白去乙酰化酶抑制剂曲古抑菌素A(trichostatinA, TSA)和帕比司他(panobinostat, LBH589)在体外对人肾癌细胞株增殖、细胞周期分布及凋亡的影响,并进一步探讨其可能的分子机制。方法LBH589或TSA作用于经或未经SP600125预处理的人肾癌细胞株OS-RC-2,在设定的时间点用四甲基偶氮唑蓝方法(MTT)检测细胞生长抑制率并检测出最佳刺激浓度和时间;用流式细胞术检测药物作用后肾癌细胞周期变化情况及凋亡情况;用Western blotting分析药物作用后肾癌细胞内c-Jun、磷酸化c-Jun(p-c-Jun)、Bcl-2、Bax蛋白的表达变化。用JNK抑制剂SP600125阻断JNK信号通路后,观察SP600125预处理+TSA组较TSA单独处理肾癌细胞周期、凋亡及凋亡相关蛋白的变化情况。结果TSA和LBH589在体外均可抑制肾癌细胞生长,且具有浓度与时间依赖性;与对照组相比,TSA或LBH589可使肾癌细胞发生G2/M期周期阻滞,并均可诱导肾癌细胞发生明显凋亡。Western blotting显示TSA和LBH589均能明显促进p-c-jun蛋白的表达,同时TSA也能诱导Bax蛋白表达而抑制Bcl2蛋白表达,与对照组相比有显著统计学意义(P<0.05);与TSA单独处理相比,JNK抑制剂SP600125预处理+TSA能部分减弱TSA对肾癌细胞的抑制效应并逆转TSA诱导的肾癌细胞G2/M期阻滞和细胞凋亡,差异有统计学意义(P<0.05)。结论组蛋白去乙酰化酶抑制剂在体外可抑制OS-RC-2细胞增殖,阻滞细胞周期、诱导细胞凋亡;TSA可通过JNK信号通路诱导OS-RC-2细胞G2/M期阻滞,调节凋亡蛋白的表达从而诱导细胞凋亡。
目的研究組蛋白去乙酰化酶抑製劑麯古抑菌素A(trichostatinA, TSA)和帕比司他(panobinostat, LBH589)在體外對人腎癌細胞株增殖、細胞週期分佈及凋亡的影響,併進一步探討其可能的分子機製。方法LBH589或TSA作用于經或未經SP600125預處理的人腎癌細胞株OS-RC-2,在設定的時間點用四甲基偶氮唑藍方法(MTT)檢測細胞生長抑製率併檢測齣最佳刺激濃度和時間;用流式細胞術檢測藥物作用後腎癌細胞週期變化情況及凋亡情況;用Western blotting分析藥物作用後腎癌細胞內c-Jun、燐痠化c-Jun(p-c-Jun)、Bcl-2、Bax蛋白的錶達變化。用JNK抑製劑SP600125阻斷JNK信號通路後,觀察SP600125預處理+TSA組較TSA單獨處理腎癌細胞週期、凋亡及凋亡相關蛋白的變化情況。結果TSA和LBH589在體外均可抑製腎癌細胞生長,且具有濃度與時間依賴性;與對照組相比,TSA或LBH589可使腎癌細胞髮生G2/M期週期阻滯,併均可誘導腎癌細胞髮生明顯凋亡。Western blotting顯示TSA和LBH589均能明顯促進p-c-jun蛋白的錶達,同時TSA也能誘導Bax蛋白錶達而抑製Bcl2蛋白錶達,與對照組相比有顯著統計學意義(P<0.05);與TSA單獨處理相比,JNK抑製劑SP600125預處理+TSA能部分減弱TSA對腎癌細胞的抑製效應併逆轉TSA誘導的腎癌細胞G2/M期阻滯和細胞凋亡,差異有統計學意義(P<0.05)。結論組蛋白去乙酰化酶抑製劑在體外可抑製OS-RC-2細胞增殖,阻滯細胞週期、誘導細胞凋亡;TSA可通過JNK信號通路誘導OS-RC-2細胞G2/M期阻滯,調節凋亡蛋白的錶達從而誘導細胞凋亡。
목적연구조단백거을선화매억제제곡고억균소A(trichostatinA, TSA)화파비사타(panobinostat, LBH589)재체외대인신암세포주증식、세포주기분포급조망적영향,병진일보탐토기가능적분자궤제。방법LBH589혹TSA작용우경혹미경SP600125예처리적인신암세포주OS-RC-2,재설정적시간점용사갑기우담서람방법(MTT)검측세포생장억제솔병검측출최가자격농도화시간;용류식세포술검측약물작용후신암세포주기변화정황급조망정황;용Western blotting분석약물작용후신암세포내c-Jun、린산화c-Jun(p-c-Jun)、Bcl-2、Bax단백적표체변화。용JNK억제제SP600125조단JNK신호통로후,관찰SP600125예처리+TSA조교TSA단독처리신암세포주기、조망급조망상관단백적변화정황。결과TSA화LBH589재체외균가억제신암세포생장,차구유농도여시간의뢰성;여대조조상비,TSA혹LBH589가사신암세포발생G2/M기주기조체,병균가유도신암세포발생명현조망。Western blotting현시TSA화LBH589균능명현촉진p-c-jun단백적표체,동시TSA야능유도Bax단백표체이억제Bcl2단백표체,여대조조상비유현저통계학의의(P<0.05);여TSA단독처리상비,JNK억제제SP600125예처리+TSA능부분감약TSA대신암세포적억제효응병역전TSA유도적신암세포G2/M기조체화세포조망,차이유통계학의의(P<0.05)。결론조단백거을선화매억제제재체외가억제OS-RC-2세포증식,조체세포주기、유도세포조망;TSA가통과JNK신호통로유도OS-RC-2세포G2/M기조체,조절조망단백적표체종이유도세포조망。
Objective To study the effect of histone deacetylase inhibitors trichostatin A (TSA) and LBH589 on the growth of human renal cell carcinoma OS-RC-2 cells in vitro and explore the underlying molecular mechanism. Methods OS-RC-2 cells were treated with LBH589 or TSA with or without SP600125 pretreatment, and the cell viability was measured by MTT assay. The changes of cell cycle distribution and apoptosis of OS-RC-2 cells were examined by flow cytometry, and the expressions of c-Jun, p-c-Jun, Bcl-2, and Bax were quantified by Western blotting. Results TSA and LBH589 both inhibited the growth of OS-RC-2 cells in a dose-and time-dependent manner. TSA at 1μnmol/L and LBH589 at 50 nmol/L caused obvious cell cycle arrest in G2/M phase and cell apoptosis, and significantly increased the protein levels of phosphorylated c-Jun. TSA treatment obviously increased Bax expression but decreased Bcl2 expression in the cells. The growth inhibitory effect of TSA was attenuated by the JNK inhibitor SP600125 in OS-RC-2 cells. TSA-induced phosphorylation of c-Jun and Bax upregulation was partially counteracted by SP600125. Conclusion TSA and LBH589 can cause cell cycle arrest and induce apoptosis in OS-RC-2 cells, in which process P-JNK pathway plays an important role.