PDTC对TRAIL诱导Raji细胞凋亡的影响
摘要
目的:探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)联合吡咯烷二硫代氨基甲酸盐(pyrrolidine dithioearbamate,PDTC)对人Burkitt淋巴瘤细胞株Raji细胞凋亡的影响,并初步探讨其作用机制。
方法:应用MTT法检测TRAIL(1、10、100ng/ml)、100μmol/mlPDTC及TRAIL(1、10、100ng/ml)+100μmol/mlPDTC时Raji细胞的生长抑制率;原位末端酶标记技术(TUNEL)检测Raji细胞凋亡指数(AI)。Western blot检测单独应用TRAIL(1、10、100ng/ml)、100μmol/mlPDTC及TRAIL(1、10、100ng/ml)联合100μmol/mlPDTC处理Raji细胞24h后,Raji细胞表达膜蛋白DR4、DR5及核内蛋白NF-κB的情况。
结果:1. 1ng/ml、10ng/ml TRAIL组12h抑制率分别为-35.52±5.64(%)及-15.07±2.03(%);100ng/mlTRAIL组12 h抑制率为6.68±1.17(%),并且呈时间依赖性(P<0.05)。100μmol/mlPDTC组12 h抑制率为1.01±0.21(%),48 h抑制率为2.12±0.33(%),无明显抑制作用。TRAIL(1、10、100ng/ml)联合100μmol/ml PDTC后,12h抑制率分别1.18±0.51、4.96±1.34、14.63±2.57(%),均显著高于相同浓度TRAIL及PDTC单用组(p<0.01),并且呈时间依赖性(P<0.05)。10ng/ml、100ng/mlTRAIL联合PDTC对Raji细胞的增殖的抑制作用均具有协同效应,其中100ng/mlTRAIL联合PDTC具有显著的协同效应。
2.联合用药组、TRAIL(100ng/ml)组及48 h 100μmol/ml PDTC凋亡指数与细胞对照组比较均有统计学意义(p<0.05)。TRAIL(100ng/ml)联合PDTC时细胞凋亡指数最高为79.49±1.40(%),较TRAIL100ng/ml(28.84±2.47%)有显著性升高, 1ng/ml、10ng/ml TRAIL联合PDTC作用后Raji凋亡细胞也均显著高于相同浓度TRAIL及PDTC单用组(p<0.01),并且呈时间依赖性。与MTT法检测结果具有一致性。
3.作用24 h后,TRAIL(1、10、100ng/ml)三组膜蛋白DR4、DR5吸光度值较细胞对照组均有统计学意义(P<0.05,p<0.01,p<0.01),而与相同浓度的TRAIL联合PDTC100μmol/ml组却无显著性差异(p>0.05)。细胞对照组核内蛋白NF-κB(p65)为0,TRAIL(1、10、100ng/ml)三组NF-κBp65吸光度值分别0.7835±0.019、0.6607±0.0310、.5194±0.024,均显著高于相同浓度的TRAIL联合PDTC组核内蛋白NF-κBp65吸光度值(0.3377±0.046、0.4837±0.012、0.6578±0.025)。100μmol/mlPDTC组膜蛋白DR4、DR5及核内蛋白NF-κBp65吸光度值均与细胞对照组相同。
结论:
1.TRAIL对Raji细胞的生长具有抑制作用,但作用不敏感。NF-κB抑制剂PDTC能显著增强TRAIL对Raji细胞的抑制作用。
2. TUNEL检测结果显示PDTC主要是通过增加肿瘤细胞凋亡来增强TRAIL对Raji细胞的抑制作用。
3.本实验证实:TRAIL通过与细胞膜上的死亡受体(DR4、DR5)结合而激活凋亡信号途径,TRAIL在激活凋亡信号途径的同时也激活了核蛋白NF-κB。PDTC通过抑制NF-κB的活化来增加Raji细胞对TRAIL诱凋亡的敏感性,死亡受体的表达不参与这一变化过程。
Objective:
To investigate whether PDTC could increase the Raji cells apoptosis induced by TRAIL and investigate the underlying mechanism.
Methods:
Using MTT assay to measure the cell growth inhibiting rate of Raji cell induced by TRAIL(1 ng/ml、10 ng/ml、100ng/ml )、PDTC(100μmol/ml)and TRAIL ( 1 ng/ml、10 ng/ml、100ng/ml ) combined with PDTC(100μmol/ml).The Raji cells apoptosis indix was examined by terminal deoxynucleotide transferas-mediated aUTP nick end labeling(TUNEL) .The expression of Membrane Proteins DR4、DR5 and nuclear protein NF-κB were detected by Western blot before and after the treatment of TRAIL、PDTC and co-treated with TRAIL and PDTC for 24h.
Results:
1. The 12h inhibiting rate of 1ng/ml、10ng/ml TRAIL was -35.52±5.64(%)and -15.07±2.03(%), the 12h inhibiting rate of 100ng/ml TRAIL was 6.68±1.17(%) and showed a time-dependent.The 12h inhibiting rate of 100μmol/mlPDTC was 1.01±0.21(%) and The 48 h inhibiting rate was 2.12±0.33(%),which showed a no significant inhibition on Raji cells.The 12 h inhibiting rate of TRAIL(1、10、100ng/ml)combined with the PDTC were 1.18±0.51、4.96±1.34、14.63±2.57(%),and were all markedly higher than in TRAIL and PDTC used alone(p<0.01).
2. The co-treated group、TRAIL(100ng/ml)group and 100μmol/ml PDTC group after 48 h were all showed significantly different to control group(p<0.05).The highest apoptosis index of Raji cells induced by TRAIL combined with the PDTC was 79.49±1.40(%) which markedly higher than TRAIL100ng/ml(28.84±2.47%). The apoptosis index of Raji cell in 1ng/ml、10ng/ml TRAIL combined with the PDTC were also markedly higher than TRAIL and PDTC used alone (P<0.01)and also showed a time-dependent, which consistent of MTT.
3.The expression of Membrane Proteins DR4、DR5 after treated with TRAIL(1、10、100ng/ml)for 24 h showed significantly diference in control group(P<0.05,p<0.01,p<0.01),but had no significantly difference in TRAIL co-treat with PDTC (p>0.05). The expression of nuclear protein NF-κB in control group is zero,but in TRAIL(1、10、100ng/ml) treated group was 0.7835±0.019、0.6607±0.0310、0.5194±0.024 which were all markedly higher than TRAIL co-treated with PDTC ( 0.3377±0.046、0.4837±0.012、0.6578±0.025). The Membrane Proteins DR4、DR5 and nuclear protein NF-κB at 100μmol/mlPDTC group were same as cotrol group.
Conclusions:
1. Raji cell is insensitive to TRAIL, PDTC could significantly increase the killing effect of TRAIL in Raji cell.
2. The TUNEL show that PDTC primarily through increasing tumor cells apoptosis to enhance the killing effect of TRAIL.
3. This study verified that TRAIL could establish a complex with death receptors and at the same time activate NF-κB.PDTC through significantly decrease the activation of NF-κB to increase the sensitiveity of TRAIL-induced apoptosis in Raji cell and the expression of death receptor not involved in this process.
方法:应用MTT法检测TRAIL(1、10、100ng/ml)、100μmol/mlPDTC及TRAIL(1、10、100ng/ml)+100μmol/mlPDTC时Raji细胞的生长抑制率;原位末端酶标记技术(TUNEL)检测Raji细胞凋亡指数(AI)。Western blot检测单独应用TRAIL(1、10、100ng/ml)、100μmol/mlPDTC及TRAIL(1、10、100ng/ml)联合100μmol/mlPDTC处理Raji细胞24h后,Raji细胞表达膜蛋白DR4、DR5及核内蛋白NF-κB的情况。
结果:1. 1ng/ml、10ng/ml TRAIL组12h抑制率分别为-35.52±5.64(%)及-15.07±2.03(%);100ng/mlTRAIL组12 h抑制率为6.68±1.17(%),并且呈时间依赖性(P<0.05)。100μmol/mlPDTC组12 h抑制率为1.01±0.21(%),48 h抑制率为2.12±0.33(%),无明显抑制作用。TRAIL(1、10、100ng/ml)联合100μmol/ml PDTC后,12h抑制率分别1.18±0.51、4.96±1.34、14.63±2.57(%),均显著高于相同浓度TRAIL及PDTC单用组(p<0.01),并且呈时间依赖性(P<0.05)。10ng/ml、100ng/mlTRAIL联合PDTC对Raji细胞的增殖的抑制作用均具有协同效应,其中100ng/mlTRAIL联合PDTC具有显著的协同效应。
2.联合用药组、TRAIL(100ng/ml)组及48 h 100μmol/ml PDTC凋亡指数与细胞对照组比较均有统计学意义(p<0.05)。TRAIL(100ng/ml)联合PDTC时细胞凋亡指数最高为79.49±1.40(%),较TRAIL100ng/ml(28.84±2.47%)有显著性升高, 1ng/ml、10ng/ml TRAIL联合PDTC作用后Raji凋亡细胞也均显著高于相同浓度TRAIL及PDTC单用组(p<0.01),并且呈时间依赖性。与MTT法检测结果具有一致性。
3.作用24 h后,TRAIL(1、10、100ng/ml)三组膜蛋白DR4、DR5吸光度值较细胞对照组均有统计学意义(P<0.05,p<0.01,p<0.01),而与相同浓度的TRAIL联合PDTC100μmol/ml组却无显著性差异(p>0.05)。细胞对照组核内蛋白NF-κB(p65)为0,TRAIL(1、10、100ng/ml)三组NF-κBp65吸光度值分别0.7835±0.019、0.6607±0.0310、.5194±0.024,均显著高于相同浓度的TRAIL联合PDTC组核内蛋白NF-κBp65吸光度值(0.3377±0.046、0.4837±0.012、0.6578±0.025)。100μmol/mlPDTC组膜蛋白DR4、DR5及核内蛋白NF-κBp65吸光度值均与细胞对照组相同。
结论:
1.TRAIL对Raji细胞的生长具有抑制作用,但作用不敏感。NF-κB抑制剂PDTC能显著增强TRAIL对Raji细胞的抑制作用。
2. TUNEL检测结果显示PDTC主要是通过增加肿瘤细胞凋亡来增强TRAIL对Raji细胞的抑制作用。
3.本实验证实:TRAIL通过与细胞膜上的死亡受体(DR4、DR5)结合而激活凋亡信号途径,TRAIL在激活凋亡信号途径的同时也激活了核蛋白NF-κB。PDTC通过抑制NF-κB的活化来增加Raji细胞对TRAIL诱凋亡的敏感性,死亡受体的表达不参与这一变化过程。
Objective:
To investigate whether PDTC could increase the Raji cells apoptosis induced by TRAIL and investigate the underlying mechanism.
Methods:
Using MTT assay to measure the cell growth inhibiting rate of Raji cell induced by TRAIL(1 ng/ml、10 ng/ml、100ng/ml )、PDTC(100μmol/ml)and TRAIL ( 1 ng/ml、10 ng/ml、100ng/ml ) combined with PDTC(100μmol/ml).The Raji cells apoptosis indix was examined by terminal deoxynucleotide transferas-mediated aUTP nick end labeling(TUNEL) .The expression of Membrane Proteins DR4、DR5 and nuclear protein NF-κB were detected by Western blot before and after the treatment of TRAIL、PDTC and co-treated with TRAIL and PDTC for 24h.
Results:
1. The 12h inhibiting rate of 1ng/ml、10ng/ml TRAIL was -35.52±5.64(%)and -15.07±2.03(%), the 12h inhibiting rate of 100ng/ml TRAIL was 6.68±1.17(%) and showed a time-dependent.The 12h inhibiting rate of 100μmol/mlPDTC was 1.01±0.21(%) and The 48 h inhibiting rate was 2.12±0.33(%),which showed a no significant inhibition on Raji cells.The 12 h inhibiting rate of TRAIL(1、10、100ng/ml)combined with the PDTC were 1.18±0.51、4.96±1.34、14.63±2.57(%),and were all markedly higher than in TRAIL and PDTC used alone(p<0.01).
2. The co-treated group、TRAIL(100ng/ml)group and 100μmol/ml PDTC group after 48 h were all showed significantly different to control group(p<0.05).The highest apoptosis index of Raji cells induced by TRAIL combined with the PDTC was 79.49±1.40(%) which markedly higher than TRAIL100ng/ml(28.84±2.47%). The apoptosis index of Raji cell in 1ng/ml、10ng/ml TRAIL combined with the PDTC were also markedly higher than TRAIL and PDTC used alone (P<0.01)and also showed a time-dependent, which consistent of MTT.
3.The expression of Membrane Proteins DR4、DR5 after treated with TRAIL(1、10、100ng/ml)for 24 h showed significantly diference in control group(P<0.05,p<0.01,p<0.01),but had no significantly difference in TRAIL co-treat with PDTC (p>0.05). The expression of nuclear protein NF-κB in control group is zero,but in TRAIL(1、10、100ng/ml) treated group was 0.7835±0.019、0.6607±0.0310、0.5194±0.024 which were all markedly higher than TRAIL co-treated with PDTC ( 0.3377±0.046、0.4837±0.012、0.6578±0.025). The Membrane Proteins DR4、DR5 and nuclear protein NF-κB at 100μmol/mlPDTC group were same as cotrol group.
Conclusions:
1. Raji cell is insensitive to TRAIL, PDTC could significantly increase the killing effect of TRAIL in Raji cell.
2. The TUNEL show that PDTC primarily through increasing tumor cells apoptosis to enhance the killing effect of TRAIL.
3. This study verified that TRAIL could establish a complex with death receptors and at the same time activate NF-κB.PDTC through significantly decrease the activation of NF-κB to increase the sensitiveity of TRAIL-induced apoptosis in Raji cell and the expression of death receptor not involved in this process.
引文
[1] Griffith TS,Anderson RD,Davidson BL,et a1.Adenoviral-mediated transfer of the TNF-related ap0pt0sis-inducing ligand/Apo-2 ligand gene induces tumor cell apoptosis[J].Immunol,2000,89:2887—2894.
[2] Sprick M R,Weigand M A,Rieser E,et a1. FADD/MORT1 and caspase一8 are recruited to TRAII receptors 1 and 2 and are essential for apoptosis mediated by TRAI1 receptor 2[J].Immunity,2000,12(6):599—609
[3] Kischkel F C,Lawrence D A,Chuntharapai A,et a1.Apo21 /TRAIL—dependent recruitment of endogenous FADD and caspase一8 to death receptors 4 and 5[J].Immunity,2000,12(6):611—62O
[4]姚悦萍,欧阳健,侯亚义,等.重组人可溶性TRAIL分子诱导白血病细胞株凋亡的研究现代免疫学,2006,26(3):234-238
[5]施敏骅,胡筠珠,胡华成. TRAIL诱导非小细胞肺癌凋亡的实验研究江苏医药杂志,2004,3O(9): 668-669
[6] Hopkins-Donaldson S,Ziegler A,Kurtz S,et a1.Silencing of death receptor and caspase-8 expression in small cell lung carcinaoma cell lines and tumors by DNA methylationl [J].ce11 Death Difer,2003,10(3):356-364
[7] Yang X,Thiele CJ.Targeting the tumor necrosis factor-related apoptosis-inducing ligand path in neuroblastoma[J].Cancer Lett,20O3,197(1-2):137—143.
[8] Evdokiou A,Bouralexis S,Atkins GJ,et a1.Chemotherapeutic agents sensitize osteogenic sarcoma cells , but not normal human bone cells , to Apo2L /TRAIL-induced apoptosis[J]. Cancer,2002,99(4):49 1—504.
[9]李娜,刁路明,邹祖玉.TRAIL与羟基喜树碱联用协同杀伤人肺腺癌A549细胞.肿瘤防治杂志,2005,12(13):961-964
[10] GilmoreTD.NF-kappa B is requiredfor H-ra8 oncogeneincduced abnomml cell proliferation and tumorigenesis [J].Oncogene,2OOO,19(7):841.849
[11] Arenzana-Seisdedos F,Turpin P,Rodriguez M,et.al.Nuclear localization of I kappa B alpha promotes axtive transport of NF kappa B from the mucleus to the cytoplasm [J]. Oncogene.1999,18(49):6867-6874. [12 ] Arenzana-Seisdedos F,Turpin P,RedrigaezM,et a1.Nuclear locallzatione of I kappa B alpha promotes active transport of NF-kappa B fromthe nucleus to the cytoplasm[J].J Cell Sci,l997,ll0(3):369—378.
[13] Shen J,Channavajhala P,Seldin DC.et al.Phosphorylation by the protein kinase CK2 promotes calpain-mediated degradation of I kappaB alpha. [J] Immunol. 2001 Nov 1;167(9):4919-25
[14] Zhang L,Fang B. Mechanisms of resistance to TRAIL-induced apoptosis incancer.[J]:Cancer Gene Ther. 2005 ;12(3):228-37
[15] Wachter T,Sprick M,Jausmann D, et al. cFLIPL inhibits tumor necrosis factor-related apoptosis-inducing ligand-mediated NF-kappaB activation at the death-inducing signaling complex in human keratinocytes.[J]; J Biol Chem. 2004 17;279(51):52824-34.
[16] Kim KM,Lee YJ. Amiloride augments TRAIL-induced apoptotic death by inhibiting phosphorylation of kinases and phosphatases associated with the P13K-Akt pathway.[J]: Oncogene. 2005 13;24(3):355-66.
[17] Chen X, Kandasamy K, Srivastara PK, et al. Differential roles of RelA (p65) and c-Rel subunits of nuclear factor kappa B in tumor necrosis factor-related apoptosis-inducing ligand signaling[J].Cancer Res 2003 1;63(5):1059-66
[18] Ou D, Wang X, Metzger DL, et al. Regulation of TNF-related apoptosis-inducing ligand-mediated death-signal pathway in human beta cells by Fas-associated death domain and nuclear factor kappaB[J].Hum Immunol. 2005;66(7):799-809.
[19] Kim HS, Chang I, Kim JY, et al. Caspase-mediated p65 cleavage promotes TRAIL-induced apoptosis[J], Cancer Res,2005 15;65(14):6111-9
[20] Oya M, Ohtsubo M, Takayanagi A, et al. Constitutive activation of nuclear factor kB prevents TRAIL- induced apoptosis in renal cancer cells[J]. Oncogene, 2001 28;20(29):3888-96
[21] Hu WH, Johnson H, Shu HB, et al. Tumor necrosis factor-related apoptosis -inducing ligand receptors signal NF-kaptpa B and JNK activation and apoptosis through distinct pathways[J] J Biol Chem,1999 22;274(43) : 30603-10.
[22] Mouzakiti A, Packham G. Regulation of tumour necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in Burkitt's lymphoma cell lines[J]. Br J Haematol. 2003 ;122(1):61-9.
[23] Tafuku S, Matsuda T, Kawakami H, et al. Potential mechanism of resistance to TRAIL-imduced apoptosis in Burkitt’s lymphoma[J].Eur J Haematol, 2006 Jan;76(1):64-74.
[24]许良中主编.现代恶性淋巴瘤病理学.上海:上海科技文献出版社(第一版);2002,39
[25] Yang X,Thiele CJ.Targeting the tumor necrosis factor-related apoptosis- inducing ligand path in neuroblastoma[J].Cancer Lett,2003,197:137_143.
[26]叶子茵,林汉良.细胞对TRAIL诱导敏感性的调控机制.临床与实验病理学杂志.2006;22(5):608-610
[27] Lauzurica P,Marazuela M,et al .Pyrrolidine dithiocarbamate protects mice from lethal shock induced by LPS or TNF-alpha[J] .Eur J Immunol ,1999,29(6):1890-1900
[28] LI Ya-qing,ZHANG Zhen-xiang,XU Yong-jian,et a1.N-Acetyl-L-cysteine an d pyrrolidine dithiocarbamate in-hibited nuclear factor-KB activation in alveolar macrophages by different mechanisms[J].Acta PharmacolSin,2006,27(3):339—346
[29] Morais C , Pat B , Gobe G , et a1 . Pyrrolidine dithiocar-bamate exerts anti-proliferative an d pro-apoptotic effectsin renal cell carcinoma cell lines[J].Nephrol Dial Transplant,2006,21(12):3377—3388
[30] Yeh Y A,Herenyiova M ,W eber G.Quercetin:synergistic action with carboxyamidotriazole in human breast carcinom a cells[J].Life Sci,1995, 57(13):1285-1292
[31] Almasan A,Ashkenazi A.Apo2L/TBAIL:apoptosis signaling,biology,and potential for cancer thempy[J].Cytokin Growth Factor,2003,14(3—4):337
[32] Pitti RM .Marsters SA,Ruppest S,et a1.Induction of aoptosis by Apo-2 1igand,a new member of the tumor necrosis factor cytoklne family[J]. Biol Chem, 1996,27l:12687-l2690
[33] Michael S,Volker R,Lorenz CH,et a1.TNF-related apoptosis-inducing ligand and its decoy receptor osteopmtegerin in nonischemic dilated cardiomyopathy[J].Biochemical and BiophysicalResearch Communications,2005,338(4):1745—1750
[34] Holen I,Croucher PI,Hamdy FC,et a1.Osteoprotegerin(OPG)is a survival factor for human prostate cancer cells[J].Cancer Res,2002,62(6):1619-1623
[35] Ashkenazi A,Pai RC,Fong S,et a1.Safety and antitumoractivityof recombinant soluble Apo2 ligand[J].J Clin Invest,1999,104:155
[36] Keane MM.Ettenberg SA,Nan MM,et a1.Chemotherapy augments TRAIL-induced apoptosis in breast cell lines[J].Cancer Res,1999.59(3):734-741.
[37] Evdokiou A,Bouralexis S,Atkins GJ,et a1.Chemotherapeutic agents sensitize osteogenic sarcoma cells , but not normal humanbone cells , to Apo2L /TRAIL-induced apoptosis[J].Int J Cancer,2002,99(4):491-504 .
[38] Hemandez A,Wang QD,Schwartz SA,et a1.Sensitization of human colon cancer cells to TRAIL-mediated apoptosis[J].J Gastrointest Surg,2001,5(1):56-65.
[39] Song JJ,An JY,Kwon YT,et a1.Evidence oftwomodes ofdevelopment of acquired TRAIL resistance.Involvement of Bcl-xL[J].J Biol Chem.2007,282(1):319-328
[40] Ehrhardt H , Fulda S , Schmed I , et al. TRAIL induced survival and proliferation in cancer cells resistant towards TRAIL-induced apoptosis mediated by NF-kappaB[J].Oncogene ,2003 ,22 :3842
[41] PanG,O’RoarkeK,ChinmiyanAM,et a1.The receptor for the cytotoxic ligand TRAIL[J]Science,1997,276(5309):111.113
[42] Shivapurdar N, Toyooka S, Toyooka KD, et al Aberranl methylation of trail decoy receptor genes is frequent in multiple tumore types[J] .Int J cancer. 2004. 109:786-792
[43] Truneh A,Sharma S,Silveman C,et a1.Temperature-sens/five diferential afinity of TRAIL for its receptors DR5 is the highest afinity receptor[J].JBiol Chem,2000,275(30):23319
[44] Karin M.Nuclear factor-kappa B in cancer development and progression[J].Nature,2006,441(7902):431—436.
[45] Mistry P,Deacon K,Mistry S,et a1.NF-KB Promotes Survival during Mitotic Cell Cycle Arrest[J].J Biol Chem,2004,279(2):1482—1490
[1] Sen R,Baltimore D, Multiple nuclear factors interact with the immunoglobulin enhancer sequences [J]: Cell, 1986 Aug 29;46(5):705-16.
[2] Zhang L,Fang B. Mechanisms of resistance to TRAIL-induced apoptosis in cancer.[J]:Cancer Gene Ther. 2005 Mar;12(3):228-37
[3] Wachter T,Sprick M,Jausmann D, et al. cFLIPL inhibits tumor necrosis factor-related apoptosis-inducing ligand-mediated NF-kappaB activation at the death-inducing signaling complex in human keratinocytes.[J]; J Biol Chem. 2004 Dec 17;279(51):52824-34.
[4]Kim KM,Lee YJ. Amiloride augments TRAIL-induced apoptotic death by inhibiting phosphorylation of kinases and phosphatases associated with the P13K-Akt pathway.[J]: Oncogene. 2005 Jan 13;24(3):355-66.
[5]Arenzana-Seisdedos F,Turpin P,Rodriguez M,et.al.Nuclear localization of I kappa B alpha promotes axtive transport of NF kappa B from the mucleus to the cytoplasm [J]. Oncogene.1999,18(49):6867-6874.
[6]Shen J,Channavajhala P,Seldin DC.et al.Phosphorylation by the protein kinase CK2 promotes calpain-mediated degradation of IkappaBalpha.J Immunol. 2001 Nov 1;167(9):4919-25
[7]Chen X, Kandasamy K, Srivastara PK, et al. Differential roles of RelA (p65) and c-Rel subunits of nuclear factor kappa B in tumor necrosis factor-related apoptosis-inducing ligand signaling.Cancer Res 2003 Mar 1; 63(5):1059-66
[8]Bernard D, Quatannens B, Vandenbunder B, et al. Rel/NF-kappaB transcription factors protect against tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-induced apoptosis by up-regulating the TRAIL decoy receptor DcR1. J Biol Chem. 2001 Jul 20;276(29):27322-8
[9]Baetu TM, Kwon H, Sharma S, et al. Disruption of NF-kappaB signaling reveals a novel role for NF-kappaB in the regulation of TNF-related apoptosis-inducing ligand expression. J Immunol. 2001 Sep 15;167(6):3164-73
[10]Shetty S, Gladden JB, Henson Es, et al. Tumor necrosisfactor-related apoptosis inducing ligand(TRAIL) upregulations death receptor 5 (DR5)mediated by NFkappaBactivation in epithelial derved cell lines[J].Apoptosis, 2002 Oct;7(5):413-20.
[11]Ou D, Wang X, Metzger DL, et al. Regulation of TNF-related apoptosis-inducing ligand-mediated death-signal pathway in human beta cells by Fas-associated death domain and nuclear factor kappaB.Hum Immunol. 2005 Jul;66(7):799-809.
[12]Kim HS, Chang I, Kim JY, et al. Caspase-mediated p65 cleavage promotes TRAIL-induced apoptosis, Cancer Res,2005 Jul 15;65(14):6111-9
[13]Mendoza FJ, Ishdori G, Hu X, et al. Death receptor-4 (DR4) expression is regulated by transcription factor NF-kappaB in response to etoposide treatment. Apoptosis. 2008 Jun;13(6):756-70.
[14]Ehrhardt H, Fulda S, Schmed I, et a1.TRAII inducedsurvival and proliferation in cancer cells resistant towardsTRAIL-induced apoptosis mediated by NF-kappaB[J].Oncogene, 2003 Jun 19;22(25):3842-52.
[15]Tafuku S, Matsuda T, Kawakami H, et al. Potential mechanism of resistance to TRAIL-imduced apoptosis in Burkitt’s lymphoma.Eur J Haematol, 2006 Jan;76(1):64-74.
[16]Hu WH, Johnson H, Shu HB, et al. Tumor necrosis factor-related apoptosis-inducing ligand receptors signal NF-kaptpa B and JNK activation and apoptosis through distinct pathways[J] J Biol Chem,1999 Oct 22;274(43):30603-10.
[17] Wajant H,Haas E,Schwenzer R,et al.Inhibition of death receptor-mediated gene induction by a cycloheximide-sensitive factor occurs at the level of or upstream lf Fas-associated death domain protein(FADD)[J] J Biol chem, 2000 Aug 11;275(32):24357-66.
[18]Karacay B, Sanlioqlu S,Griffith TS, et al. Inhibition of the NF-kappaB pathway enhances TRAIL-mediated apoptosis in neuroblastoma cells.Cancer Gene Ther. 2004 Oct;11(10):681-90
[19]朱华云,刘凌翔,葛红梅等,NF-kB抑制剂PDTC对人胃癌SGC790l细胞凋亡及Livin和Caspase-3表达的影响[J]南京医科大学学报,2007,27(8)816-820.
[20]Frelin C, Imbert V, Griessinger E, et al.Targeting NF-kB kinase activity via pharmacological inhibition of IKK2-induced apoptosis of human acute myeloid leukemia cells[J].Blood, 2005 Jan 15;105(2):804-11.
[21]Richardson PG, Barlogie B, Berenson J, et a1. A phase 2 study ofbortezomib in relap sed refractorymyeloma. N Engl J Med. 2003 Jun 26;348(26):2609-17.
[22]俞呜,李强,郝继辉等,核转录因子NF-kB在TRAIL诱导肝癌细胞凋亡中作用的研究.[J]中华肝胆外科杂志,2006.12(12)850-852.
[2] Sprick M R,Weigand M A,Rieser E,et a1. FADD/MORT1 and caspase一8 are recruited to TRAII receptors 1 and 2 and are essential for apoptosis mediated by TRAI1 receptor 2[J].Immunity,2000,12(6):599—609
[3] Kischkel F C,Lawrence D A,Chuntharapai A,et a1.Apo21 /TRAIL—dependent recruitment of endogenous FADD and caspase一8 to death receptors 4 and 5[J].Immunity,2000,12(6):611—62O
[4]姚悦萍,欧阳健,侯亚义,等.重组人可溶性TRAIL分子诱导白血病细胞株凋亡的研究现代免疫学,2006,26(3):234-238
[5]施敏骅,胡筠珠,胡华成. TRAIL诱导非小细胞肺癌凋亡的实验研究江苏医药杂志,2004,3O(9): 668-669
[6] Hopkins-Donaldson S,Ziegler A,Kurtz S,et a1.Silencing of death receptor and caspase-8 expression in small cell lung carcinaoma cell lines and tumors by DNA methylationl [J].ce11 Death Difer,2003,10(3):356-364
[7] Yang X,Thiele CJ.Targeting the tumor necrosis factor-related apoptosis-inducing ligand path in neuroblastoma[J].Cancer Lett,20O3,197(1-2):137—143.
[8] Evdokiou A,Bouralexis S,Atkins GJ,et a1.Chemotherapeutic agents sensitize osteogenic sarcoma cells , but not normal human bone cells , to Apo2L /TRAIL-induced apoptosis[J]. Cancer,2002,99(4):49 1—504.
[9]李娜,刁路明,邹祖玉.TRAIL与羟基喜树碱联用协同杀伤人肺腺癌A549细胞.肿瘤防治杂志,2005,12(13):961-964
[10] GilmoreTD.NF-kappa B is requiredfor H-ra8 oncogeneincduced abnomml cell proliferation and tumorigenesis [J].Oncogene,2OOO,19(7):841.849
[11] Arenzana-Seisdedos F,Turpin P,Rodriguez M,et.al.Nuclear localization of I kappa B alpha promotes axtive transport of NF kappa B from the mucleus to the cytoplasm [J]. Oncogene.1999,18(49):6867-6874. [12 ] Arenzana-Seisdedos F,Turpin P,RedrigaezM,et a1.Nuclear locallzatione of I kappa B alpha promotes active transport of NF-kappa B fromthe nucleus to the cytoplasm[J].J Cell Sci,l997,ll0(3):369—378.
[13] Shen J,Channavajhala P,Seldin DC.et al.Phosphorylation by the protein kinase CK2 promotes calpain-mediated degradation of I kappaB alpha. [J] Immunol. 2001 Nov 1;167(9):4919-25
[14] Zhang L,Fang B. Mechanisms of resistance to TRAIL-induced apoptosis incancer.[J]:Cancer Gene Ther. 2005 ;12(3):228-37
[15] Wachter T,Sprick M,Jausmann D, et al. cFLIPL inhibits tumor necrosis factor-related apoptosis-inducing ligand-mediated NF-kappaB activation at the death-inducing signaling complex in human keratinocytes.[J]; J Biol Chem. 2004 17;279(51):52824-34.
[16] Kim KM,Lee YJ. Amiloride augments TRAIL-induced apoptotic death by inhibiting phosphorylation of kinases and phosphatases associated with the P13K-Akt pathway.[J]: Oncogene. 2005 13;24(3):355-66.
[17] Chen X, Kandasamy K, Srivastara PK, et al. Differential roles of RelA (p65) and c-Rel subunits of nuclear factor kappa B in tumor necrosis factor-related apoptosis-inducing ligand signaling[J].Cancer Res 2003 1;63(5):1059-66
[18] Ou D, Wang X, Metzger DL, et al. Regulation of TNF-related apoptosis-inducing ligand-mediated death-signal pathway in human beta cells by Fas-associated death domain and nuclear factor kappaB[J].Hum Immunol. 2005;66(7):799-809.
[19] Kim HS, Chang I, Kim JY, et al. Caspase-mediated p65 cleavage promotes TRAIL-induced apoptosis[J], Cancer Res,2005 15;65(14):6111-9
[20] Oya M, Ohtsubo M, Takayanagi A, et al. Constitutive activation of nuclear factor kB prevents TRAIL- induced apoptosis in renal cancer cells[J]. Oncogene, 2001 28;20(29):3888-96
[21] Hu WH, Johnson H, Shu HB, et al. Tumor necrosis factor-related apoptosis -inducing ligand receptors signal NF-kaptpa B and JNK activation and apoptosis through distinct pathways[J] J Biol Chem,1999 22;274(43) : 30603-10.
[22] Mouzakiti A, Packham G. Regulation of tumour necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in Burkitt's lymphoma cell lines[J]. Br J Haematol. 2003 ;122(1):61-9.
[23] Tafuku S, Matsuda T, Kawakami H, et al. Potential mechanism of resistance to TRAIL-imduced apoptosis in Burkitt’s lymphoma[J].Eur J Haematol, 2006 Jan;76(1):64-74.
[24]许良中主编.现代恶性淋巴瘤病理学.上海:上海科技文献出版社(第一版);2002,39
[25] Yang X,Thiele CJ.Targeting the tumor necrosis factor-related apoptosis- inducing ligand path in neuroblastoma[J].Cancer Lett,2003,197:137_143.
[26]叶子茵,林汉良.细胞对TRAIL诱导敏感性的调控机制.临床与实验病理学杂志.2006;22(5):608-610
[27] Lauzurica P,Marazuela M,et al .Pyrrolidine dithiocarbamate protects mice from lethal shock induced by LPS or TNF-alpha[J] .Eur J Immunol ,1999,29(6):1890-1900
[28] LI Ya-qing,ZHANG Zhen-xiang,XU Yong-jian,et a1.N-Acetyl-L-cysteine an d pyrrolidine dithiocarbamate in-hibited nuclear factor-KB activation in alveolar macrophages by different mechanisms[J].Acta PharmacolSin,2006,27(3):339—346
[29] Morais C , Pat B , Gobe G , et a1 . Pyrrolidine dithiocar-bamate exerts anti-proliferative an d pro-apoptotic effectsin renal cell carcinoma cell lines[J].Nephrol Dial Transplant,2006,21(12):3377—3388
[30] Yeh Y A,Herenyiova M ,W eber G.Quercetin:synergistic action with carboxyamidotriazole in human breast carcinom a cells[J].Life Sci,1995, 57(13):1285-1292
[31] Almasan A,Ashkenazi A.Apo2L/TBAIL:apoptosis signaling,biology,and potential for cancer thempy[J].Cytokin Growth Factor,2003,14(3—4):337
[32] Pitti RM .Marsters SA,Ruppest S,et a1.Induction of aoptosis by Apo-2 1igand,a new member of the tumor necrosis factor cytoklne family[J]. Biol Chem, 1996,27l:12687-l2690
[33] Michael S,Volker R,Lorenz CH,et a1.TNF-related apoptosis-inducing ligand and its decoy receptor osteopmtegerin in nonischemic dilated cardiomyopathy[J].Biochemical and BiophysicalResearch Communications,2005,338(4):1745—1750
[34] Holen I,Croucher PI,Hamdy FC,et a1.Osteoprotegerin(OPG)is a survival factor for human prostate cancer cells[J].Cancer Res,2002,62(6):1619-1623
[35] Ashkenazi A,Pai RC,Fong S,et a1.Safety and antitumoractivityof recombinant soluble Apo2 ligand[J].J Clin Invest,1999,104:155
[36] Keane MM.Ettenberg SA,Nan MM,et a1.Chemotherapy augments TRAIL-induced apoptosis in breast cell lines[J].Cancer Res,1999.59(3):734-741.
[37] Evdokiou A,Bouralexis S,Atkins GJ,et a1.Chemotherapeutic agents sensitize osteogenic sarcoma cells , but not normal humanbone cells , to Apo2L /TRAIL-induced apoptosis[J].Int J Cancer,2002,99(4):491-504 .
[38] Hemandez A,Wang QD,Schwartz SA,et a1.Sensitization of human colon cancer cells to TRAIL-mediated apoptosis[J].J Gastrointest Surg,2001,5(1):56-65.
[39] Song JJ,An JY,Kwon YT,et a1.Evidence oftwomodes ofdevelopment of acquired TRAIL resistance.Involvement of Bcl-xL[J].J Biol Chem.2007,282(1):319-328
[40] Ehrhardt H , Fulda S , Schmed I , et al. TRAIL induced survival and proliferation in cancer cells resistant towards TRAIL-induced apoptosis mediated by NF-kappaB[J].Oncogene ,2003 ,22 :3842
[41] PanG,O’RoarkeK,ChinmiyanAM,et a1.The receptor for the cytotoxic ligand TRAIL[J]Science,1997,276(5309):111.113
[42] Shivapurdar N, Toyooka S, Toyooka KD, et al Aberranl methylation of trail decoy receptor genes is frequent in multiple tumore types[J] .Int J cancer. 2004. 109:786-792
[43] Truneh A,Sharma S,Silveman C,et a1.Temperature-sens/five diferential afinity of TRAIL for its receptors DR5 is the highest afinity receptor[J].JBiol Chem,2000,275(30):23319
[44] Karin M.Nuclear factor-kappa B in cancer development and progression[J].Nature,2006,441(7902):431—436.
[45] Mistry P,Deacon K,Mistry S,et a1.NF-KB Promotes Survival during Mitotic Cell Cycle Arrest[J].J Biol Chem,2004,279(2):1482—1490
[1] Sen R,Baltimore D, Multiple nuclear factors interact with the immunoglobulin enhancer sequences [J]: Cell, 1986 Aug 29;46(5):705-16.
[2] Zhang L,Fang B. Mechanisms of resistance to TRAIL-induced apoptosis in cancer.[J]:Cancer Gene Ther. 2005 Mar;12(3):228-37
[3] Wachter T,Sprick M,Jausmann D, et al. cFLIPL inhibits tumor necrosis factor-related apoptosis-inducing ligand-mediated NF-kappaB activation at the death-inducing signaling complex in human keratinocytes.[J]; J Biol Chem. 2004 Dec 17;279(51):52824-34.
[4]Kim KM,Lee YJ. Amiloride augments TRAIL-induced apoptotic death by inhibiting phosphorylation of kinases and phosphatases associated with the P13K-Akt pathway.[J]: Oncogene. 2005 Jan 13;24(3):355-66.
[5]Arenzana-Seisdedos F,Turpin P,Rodriguez M,et.al.Nuclear localization of I kappa B alpha promotes axtive transport of NF kappa B from the mucleus to the cytoplasm [J]. Oncogene.1999,18(49):6867-6874.
[6]Shen J,Channavajhala P,Seldin DC.et al.Phosphorylation by the protein kinase CK2 promotes calpain-mediated degradation of IkappaBalpha.J Immunol. 2001 Nov 1;167(9):4919-25
[7]Chen X, Kandasamy K, Srivastara PK, et al. Differential roles of RelA (p65) and c-Rel subunits of nuclear factor kappa B in tumor necrosis factor-related apoptosis-inducing ligand signaling.Cancer Res 2003 Mar 1; 63(5):1059-66
[8]Bernard D, Quatannens B, Vandenbunder B, et al. Rel/NF-kappaB transcription factors protect against tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-induced apoptosis by up-regulating the TRAIL decoy receptor DcR1. J Biol Chem. 2001 Jul 20;276(29):27322-8
[9]Baetu TM, Kwon H, Sharma S, et al. Disruption of NF-kappaB signaling reveals a novel role for NF-kappaB in the regulation of TNF-related apoptosis-inducing ligand expression. J Immunol. 2001 Sep 15;167(6):3164-73
[10]Shetty S, Gladden JB, Henson Es, et al. Tumor necrosisfactor-related apoptosis inducing ligand(TRAIL) upregulations death receptor 5 (DR5)mediated by NFkappaBactivation in epithelial derved cell lines[J].Apoptosis, 2002 Oct;7(5):413-20.
[11]Ou D, Wang X, Metzger DL, et al. Regulation of TNF-related apoptosis-inducing ligand-mediated death-signal pathway in human beta cells by Fas-associated death domain and nuclear factor kappaB.Hum Immunol. 2005 Jul;66(7):799-809.
[12]Kim HS, Chang I, Kim JY, et al. Caspase-mediated p65 cleavage promotes TRAIL-induced apoptosis, Cancer Res,2005 Jul 15;65(14):6111-9
[13]Mendoza FJ, Ishdori G, Hu X, et al. Death receptor-4 (DR4) expression is regulated by transcription factor NF-kappaB in response to etoposide treatment. Apoptosis. 2008 Jun;13(6):756-70.
[14]Ehrhardt H, Fulda S, Schmed I, et a1.TRAII inducedsurvival and proliferation in cancer cells resistant towardsTRAIL-induced apoptosis mediated by NF-kappaB[J].Oncogene, 2003 Jun 19;22(25):3842-52.
[15]Tafuku S, Matsuda T, Kawakami H, et al. Potential mechanism of resistance to TRAIL-imduced apoptosis in Burkitt’s lymphoma.Eur J Haematol, 2006 Jan;76(1):64-74.
[16]Hu WH, Johnson H, Shu HB, et al. Tumor necrosis factor-related apoptosis-inducing ligand receptors signal NF-kaptpa B and JNK activation and apoptosis through distinct pathways[J] J Biol Chem,1999 Oct 22;274(43):30603-10.
[17] Wajant H,Haas E,Schwenzer R,et al.Inhibition of death receptor-mediated gene induction by a cycloheximide-sensitive factor occurs at the level of or upstream lf Fas-associated death domain protein(FADD)[J] J Biol chem, 2000 Aug 11;275(32):24357-66.
[18]Karacay B, Sanlioqlu S,Griffith TS, et al. Inhibition of the NF-kappaB pathway enhances TRAIL-mediated apoptosis in neuroblastoma cells.Cancer Gene Ther. 2004 Oct;11(10):681-90
[19]朱华云,刘凌翔,葛红梅等,NF-kB抑制剂PDTC对人胃癌SGC790l细胞凋亡及Livin和Caspase-3表达的影响[J]南京医科大学学报,2007,27(8)816-820.
[20]Frelin C, Imbert V, Griessinger E, et al.Targeting NF-kB kinase activity via pharmacological inhibition of IKK2-induced apoptosis of human acute myeloid leukemia cells[J].Blood, 2005 Jan 15;105(2):804-11.
[21]Richardson PG, Barlogie B, Berenson J, et a1. A phase 2 study ofbortezomib in relap sed refractorymyeloma. N Engl J Med. 2003 Jun 26;348(26):2609-17.
[22]俞呜,李强,郝继辉等,核转录因子NF-kB在TRAIL诱导肝癌细胞凋亡中作用的研究.[J]中华肝胆外科杂志,2006.12(12)850-852.