中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
1期
81-83
,共3页
王广义%邱伟%孙晓东%吕国悦
王廣義%邱偉%孫曉東%呂國悅
왕엄의%구위%손효동%려국열
癌,肝细胞%肿瘤坏死因子相关的凋亡诱导配体%受体相互作用蛋白%肿瘤坏死因子-α诱导蛋白3
癌,肝細胞%腫瘤壞死因子相關的凋亡誘導配體%受體相互作用蛋白%腫瘤壞死因子-α誘導蛋白3
암,간세포%종류배사인자상관적조망유도배체%수체상호작용단백%종류배사인자-α유도단백3
Carcinoma,hepatocellular%Tumor necrosis factor-related apoptosis-inducing ligand%Receptor-interacting protein%Tumor necrosis factor-α inducing protein 3
目的 探讨肿瘤坏死因子-α诱导蛋白3(A20)及受体作用蛋白(RIP)沉默增加肝癌细胞HepG2对肿瘤坏死因子相关的凋亡诱导配体(TRAIL)敏感性的分子机制.方法 合成针对A20基因和RIP基因的特异性小干扰RNA(siRNA)并转染至肝癌细胞HepG2中,应用酸性磷酸酶(AP)法检测A20-siRNA或RIP-siRNA联合TRAIL对肝癌细胞HepG2生长的抑制作用以及Westernblot检测对半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-8蛋白表达的影响.结果 成功合成A20-siRNA和RIP-siRNA并转染入HepG2细胞中.A20-siRNA与TRAIL联合处理HepG2细胞可活化RIP、Caspase-8和Caspase-3,诱导细胞凋亡.RIP-siRNA联合TRAIL可以明显抑制HepG2细胞的增殖(P＜0.05).另外RIP-siRNA联合TRAIL作用HepG2后可活化Caspase-8.结论 A20和RIP的表达是肝癌细胞对TRAIL耐受的主要原因,A20-siRNA或RIP-siRNA可以有效抑制肝癌HepG2细胞A20及RIP基因的表达,并增加TRAIL对肝癌HepG2细胞凋亡的诱导作用；其机制是通过“A20-RIP-Caspase-8”反应链序列调控活化下游Caspase-8,活化诱导的死亡受体传导通路来实现的.
目的 探討腫瘤壞死因子-α誘導蛋白3(A20)及受體作用蛋白(RIP)沉默增加肝癌細胞HepG2對腫瘤壞死因子相關的凋亡誘導配體(TRAIL)敏感性的分子機製.方法 閤成針對A20基因和RIP基因的特異性小榦擾RNA(siRNA)併轉染至肝癌細胞HepG2中,應用痠性燐痠酶(AP)法檢測A20-siRNA或RIP-siRNA聯閤TRAIL對肝癌細胞HepG2生長的抑製作用以及Westernblot檢測對半胱氨酰天鼕氨痠特異性蛋白酶(Caspase)-8蛋白錶達的影響.結果 成功閤成A20-siRNA和RIP-siRNA併轉染入HepG2細胞中.A20-siRNA與TRAIL聯閤處理HepG2細胞可活化RIP、Caspase-8和Caspase-3,誘導細胞凋亡.RIP-siRNA聯閤TRAIL可以明顯抑製HepG2細胞的增殖(P＜0.05).另外RIP-siRNA聯閤TRAIL作用HepG2後可活化Caspase-8.結論 A20和RIP的錶達是肝癌細胞對TRAIL耐受的主要原因,A20-siRNA或RIP-siRNA可以有效抑製肝癌HepG2細胞A20及RIP基因的錶達,併增加TRAIL對肝癌HepG2細胞凋亡的誘導作用；其機製是通過“A20-RIP-Caspase-8”反應鏈序列調控活化下遊Caspase-8,活化誘導的死亡受體傳導通路來實現的.
목적 탐토종류배사인자-α유도단백3(A20)급수체작용단백(RIP)침묵증가간암세포HepG2대종류배사인자상관적조망유도배체(TRAIL)민감성적분자궤제.방법 합성침대A20기인화RIP기인적특이성소간우RNA(siRNA)병전염지간암세포HepG2중,응용산성린산매(AP)법검측A20-siRNA혹RIP-siRNA연합TRAIL대간암세포HepG2생장적억제작용이급Westernblot검측대반광안선천동안산특이성단백매(Caspase)-8단백표체적영향.결과 성공합성A20-siRNA화RIP-siRNA병전염입HepG2세포중.A20-siRNA여TRAIL연합처리HepG2세포가활화RIP、Caspase-8화Caspase-3,유도세포조망.RIP-siRNA연합TRAIL가이명현억제HepG2세포적증식(P＜0.05).령외RIP-siRNA연합TRAIL작용HepG2후가활화Caspase-8.결론 A20화RIP적표체시간암세포대TRAIL내수적주요원인,A20-siRNA혹RIP-siRNA가이유효억제간암HepG2세포A20급RIP기인적표체,병증가TRAIL대간암HepG2세포조망적유도작용；기궤제시통과“A20-RIP-Caspase-8”반응련서렬조공활화하유Caspase-8,활화유도적사망수체전도통로래실현적.
Objective To investigate the molecular mechanism of silencing Tumor necrosis factorα inducing protein 3 (A20) and receptor-interacting protein (RIP) gene promoting tumor necrosis factorrelated apoptosis-inducing ligand (TRAIL)-induced apoptosis in hepatocellular carcinoma cell lines.Methods Synthesized A20-small interfering RNA (siRNA) and RIP-siRNA were transfected into HepG2 cells.The cell death of HepG2 was determined by acid phosphatase assay and the expression of Caspase-8 by Western blotting after A20-siRNA or RIP-siRNA combined with TRAIL treatment.Results A20-siRNA could activate RIP,Caspase-8,Caspase-3 and increase the sensitivity of HepG2 cells to TRAIL-induced apoptosis.The proliferation of HepG2 was inhibited by RIP-siRNA combined with TRAIL treatment (P ＜0.05).Furthermore,Caspase-8 was activated after RIP-siRNA combined with TRAIL treatment.Conclusion A20-RIP-Caspase-8 complex could sensitize HepG2 to TRAIL-induced apoptosis through elimination of Caspase-8 inhibition.
