中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
11期
2215-2218
,共4页
李伟%陈政%龚斐然%苗毅%徐泽宽%陶敏
李偉%陳政%龔斐然%苗毅%徐澤寬%陶敏
리위%진정%공비연%묘의%서택관%도민
胰腺癌%蛋白磷酸酶2A%斑蝥素%核因子-κB
胰腺癌%蛋白燐痠酶2A%斑蝥素%覈因子-κB
이선암%단백린산매2A%반모소%핵인자-κB
Pancreatic cancer%Protein phosphatase 2A%Cantharidin%Nuclear factor-κB
目的 探讨蛋白磷酸酶2A(PP2A)抑制剂通过激活核因子-κB(NF-κB)通路诱导胰腺癌细胞株人胰腺癌细胞(PANC-1)凋亡的机制.方法 荧光素酶报告基因检测NF-κB通路的激活水平,试剂盒检测半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-8、9活性,逆转录-聚合酶链反应(RT-PCR)检测NF-κB通路下游凋亡相关基因表达水平.结果 PP2A抑制剂斑蝥素、冈田酸可激活NF-κB通路,分别使NF-κB转录活性上升(10.11 ±4.09)倍、(16.21 ±5.75)倍.NF-κB通路抑制剂Bay 11-7082预处理,可分别使斑蝥素、冈田酸诱导的NF-κB转录活性上升幅度下降(61.19±6.08)％、(62.09±12.38)％；斑蝥素、风田酸可激活外源性凋亡通路,分别使Caspase-8活性上升(0.55 ±0.12)倍、(0.85±0.21)倍.Bay 11-7082预处理,可分别使斑蝥素、冈田酸诱导的Caspase-8活性上升幅度下降(20.99±7.13)％、(29.07±7.98)％；斑蝥素、冈田酸可激活内源性凋亡通路,分别使Caspase-9活性上升(1.35 ±0.20)倍、(1.18±0.19)倍,但Bay 11-7082预处理,对斑蝥素、冈田酸诱导的Caspase-9活性上升幅度无明显影响；斑蝥素、冈田酸可上调促凋亡基因的表达；Bay 11-7082预处理可抑制斑蝥素、冈田酸诱导的促凋亡基因表达上调.结论 PP2A抑制剂通过NF-κB通路依赖性机制激活胰腺癌细胞外源性凋亡通路,并上调促凋亡基因的表达.
目的 探討蛋白燐痠酶2A(PP2A)抑製劑通過激活覈因子-κB(NF-κB)通路誘導胰腺癌細胞株人胰腺癌細胞(PANC-1)凋亡的機製.方法 熒光素酶報告基因檢測NF-κB通路的激活水平,試劑盒檢測半胱氨酰天鼕氨痠特異性蛋白酶(Caspase)-8、9活性,逆轉錄-聚閤酶鏈反應(RT-PCR)檢測NF-κB通路下遊凋亡相關基因錶達水平.結果 PP2A抑製劑斑蝥素、岡田痠可激活NF-κB通路,分彆使NF-κB轉錄活性上升(10.11 ±4.09)倍、(16.21 ±5.75)倍.NF-κB通路抑製劑Bay 11-7082預處理,可分彆使斑蝥素、岡田痠誘導的NF-κB轉錄活性上升幅度下降(61.19±6.08)％、(62.09±12.38)％；斑蝥素、風田痠可激活外源性凋亡通路,分彆使Caspase-8活性上升(0.55 ±0.12)倍、(0.85±0.21)倍.Bay 11-7082預處理,可分彆使斑蝥素、岡田痠誘導的Caspase-8活性上升幅度下降(20.99±7.13)％、(29.07±7.98)％；斑蝥素、岡田痠可激活內源性凋亡通路,分彆使Caspase-9活性上升(1.35 ±0.20)倍、(1.18±0.19)倍,但Bay 11-7082預處理,對斑蝥素、岡田痠誘導的Caspase-9活性上升幅度無明顯影響；斑蝥素、岡田痠可上調促凋亡基因的錶達；Bay 11-7082預處理可抑製斑蝥素、岡田痠誘導的促凋亡基因錶達上調.結論 PP2A抑製劑通過NF-κB通路依賴性機製激活胰腺癌細胞外源性凋亡通路,併上調促凋亡基因的錶達.
목적 탐토단백린산매2A(PP2A)억제제통과격활핵인자-κB(NF-κB)통로유도이선암세포주인이선암세포(PANC-1)조망적궤제.방법 형광소매보고기인검측NF-κB통로적격활수평,시제합검측반광안선천동안산특이성단백매(Caspase)-8、9활성,역전록-취합매련반응(RT-PCR)검측NF-κB통로하유조망상관기인표체수평.결과 PP2A억제제반모소、강전산가격활NF-κB통로,분별사NF-κB전록활성상승(10.11 ±4.09)배、(16.21 ±5.75)배.NF-κB통로억제제Bay 11-7082예처리,가분별사반모소、강전산유도적NF-κB전록활성상승폭도하강(61.19±6.08)％、(62.09±12.38)％；반모소、풍전산가격활외원성조망통로,분별사Caspase-8활성상승(0.55 ±0.12)배、(0.85±0.21)배.Bay 11-7082예처리,가분별사반모소、강전산유도적Caspase-8활성상승폭도하강(20.99±7.13)％、(29.07±7.98)％；반모소、강전산가격활내원성조망통로,분별사Caspase-9활성상승(1.35 ±0.20)배、(1.18±0.19)배,단Bay 11-7082예처리,대반모소、강전산유도적Caspase-9활성상승폭도무명현영향；반모소、강전산가상조촉조망기인적표체；Bay 11-7082예처리가억제반모소、강전산유도적촉조망기인표체상조.결론 PP2A억제제통과NF-κB통로의뢰성궤제격활이선암세포외원성조망통로,병상조촉조망기인적표체.
Objective To investigate the apoptosis induction effect of protein phosphatase 2A (PP2A) inhibitors on human pancreatic cancer cell line (PANC-1).Methods Activity of nuclear factor-κB (NF-κB) pathway was tested by using luciferase reporter gene assay.Activities of Caspase 8 and 9 were determined by kits.The expression levels of genes downstream of the NF-κB pathway were tested by using reverse transcription-polymerase chain reaction (RT-PCR).Results Treatment with PP2A inhibitors,cantharidin and Okadaic acid,up-regulated the transcriptional activity of NF-κB by the folds of 10.11 ± 4.09,and 16.21 ± 5.75 respectively.Pretreatment with the NF-κB pathway inhibitor,Bay 11-7082,repressed the up-regulation of NF-κB transcriptional activity by (61.19 ±6.08)％ and (62.09 ± 12.38)％ respectively.Treatment with cantharidin or Okadaic acid activated extrinsic apoptosis pathway,and up-regulated the activity of Caspase 8 by the folds of 0.55 ±0.12,and 0.85 ±0.21 respectively.Pretreatment with Bay 11-7082 repressed the up-regulation of Caspase 8 activity by (20.99 ±7.13)％ and (29.07 ±7.98)％ respectively.Treatment with cantharidin or Okadaic acid also activated intrinsic apoptosis pathway,and up-regulated the activity of Caspase 9 by the folds of 1.35 ±0.20,and 1.18 ±0.19 respectively.However,pretreatment with Bay 11-7082 presented no significant effect on the up-regulation of Caspase 9 activity.Treatment with cantharidin or Okadaic acid up-regulated the expression of pro-apoptotic genes which could be repressed by the pretreatment with Bay 11-7082.Conclusion PP2A inhibitors triggered extrinsic apoptosis and up-regulated the expressions of pro-apoptotic genes in pancreatic cancer cells through NF-κB dependent pathway.
