摘要
第一部分表达Livin的逆转录病毒载体的构建、病毒包装和产生
目的：为了观察大鼠心肌livin基因的过表达对大鼠心肌细胞缺血再灌注后心肌细胞凋亡的影响，构建表达Livin的逆转录病毒载体，包装产生逆转录病毒颗粒。
方法：(1)从鼠乳腺癌细胞MA782中提取RNA，RT-PCR扩增得到Livin cDNA．先克隆入T载体(pGEM-T)，经XhoⅠ/HindⅢ双酶切和PCR扩增鉴定，得到重组子pGEM-T-Livin,进行DNA序列测定。XhoⅠ／HindⅢ双酶切下livin目的基因片段，重组入(插入)逆转录病毒载体pLNCX_2，XhoⅠ／HindⅢ双酶切和PCR扩增鉴定得到重组子pLNCX_2-Livin，将pLNCX_2-Livin转染入PT67细胞中进行病毒包装，纯化得到表达Livin的逆转录病毒颗粒。
结果：RT-PCR扩增得到816bp的Livin cDNA目的基因片段：筛选得到重组子pGEM-T-Livin，DNA序列分析证实插入序列与GenBank中XM-914797 Livin基因序列完全一致；筛选得到重组子pLNCX_2-Livin；包装得到表达Livin的逆转录病毒颗粒，滴度为3．9×10~7IU／ml。
结论：成功构建出表达Livin的逆转录病毒表达载体pLNCX_2-Livin，并包装纯化得到逆转录病毒颗粒，为下游实验奠定实验基础。
第二部分Livin对大鼠心肌缺血再灌注损伤的影响
目的：近年来研究发现凋亡参与了心肌缺血再灌注损伤的发生和发展，天冬氨酸特异的半胱氨酸蛋白酶-3(Caspase-3)被认为是凋亡发生的关键酶和标志酶，Livin是近年来发现的凋亡抑制蛋白家族中的新成员，能够通过抑制Caspase-3而发挥抑制凋亡作用，但是正常心肌细胞内Livin表达较少。为此，本实验拟转染表达livin的逆转录病毒载体，并建立大鼠心肌缺血再灌注模型，观察大鼠心肌缺血再灌注过程中Livin、Caspase-3的表达变化和二者的相关性，以及Livin对心肌梗死范围和心肌细胞凋亡的影响。
方法：健康SD大鼠90只，随机分为缺血再灌注组(IR组)、Livin组和假手术对照组(Control组)。缺血再灌注组结扎LAD缺血30min，再灌注120 min；Livin组为缺血再灌注前24h心肌内注射携带Livin的逆转录病毒载体；对照组只穿线而不结扎LAD。再灌注120 min后TTC染色测定梗死范围，硫磺素染色法测定无复流面积，SP免疫组化测定Caspase-3、Livin蛋白表达量，TUNEL法测定凋亡心肌细胞。
结果：1．免疫组化法检测心肌细胞有Livin的表达，缺血再灌注增加Livin的表达量(3．10±0．52)，但与Control组(2．35±0．47)比较无统计学差异。
2．缺血再灌注过程中Caspase-3表达增多明显，IR组(103．39±8．24)较Control组(91．39±4．82)显著增加；注射Livin后则Caspase-3的表达减少，Livin组(97．43±11．15)与IR组比较差异显著。
3．Livin组心肌梗死范围(12．80±1．19％)较IR组(20．82±2．82％)显著减少。
4．Livin组无复流面积(19．39±6．25％)较IR组(25．93±6．45％)显著缩小；
5．TUNEL法检测显示缺血再灌注过程中可见心肌细胞凋亡的发生，IR组(10．35±3．34)较Control组(1．10±0．42)显著增加；注射Livin后则细胞凋亡的发生明显减少，Livin组(1．70±1．57)与IR组比较差异显著，与Control组比较无显著差异。
结论：
1．正常大鼠心肌细胞有Livin的表达。
2．Livin可以抑制缺血再灌注心肌细胞Caspase-3的表达。
3．Livin可以减少缺血再灌注心肌梗死范围和无复流面积。
4．Livin可以抑制缺血再灌注心肌细胞凋亡的发生。
第三部分荧光定量PCR检测鼠心肌caspase-3，Livin mRNA
目的：观察大鼠心肌livin mRNA表达水平以及转染表达Livin的逆转录病毒载体后Livin在心肌表达的变化情况、缺血再灌注过程中Livin，Caspase-3的mRNA表达水平，以及Livin对Caspase-3 mRNA表达水平的影响．
方法：提取第二部分实验各组组织总RNA，逆转录为cDNA，用荧光定量PCR方法扩增检测Livin和Caspase-3 mRNA表达水平。
结果：荧光定量PCR检测到大鼠心肌细胞有Livin表达，正常组Livin mRNA表达水平(8．41±1．39)×10~(-3)，空载体组为(7．45±2．51)×10~(-3)，IR组为(7．82±3．22)×10~(-3)；Livin组为(145±89)×10~(-3))，显著高于其他三组(P<0．01)。正常组Caspase-3mRNA相对表达量(5．12±2．11)×10~(-3)；空载体组为(6．88±2．31)×10~(-3)；缺血再灌注过程中Caspase-3表达明显增加((92．1±34．6)×10~(-3)，显著高于正常对照组和空载体组(P<0．01)，说明IR可造成Caspase-3升高；Livin组Caspase-3 mRNA表达水平为(56．2±21．1)×10~(-3)，与IR组相比，显著降低(P<0．05)。说明Livin过表达可有效降低IR组细胞Caspase-3表达。
结论：1．正常大鼠心肌细胞有Livin mRNA的表达；
2．表达Livin的逆转录病毒载体可以成功转染大鼠心肌；
3．转染Livin可以降低大鼠缺血再灌注心肌细胞Caspase-3 mRNA的表达水平，有助于IR中的抗凋亡。
PartⅠConstruction and production of retroviral vectorexpressing Livin
Objectives: Construction a retroviral vector carrying Livin cDNA. To explore the effect of Livin over-expression on cardiac ischemia reperfusion.
Methods: Livin gene from mice MA782 cell was amplificated by reverse-transcription polymerase chain reaction (RT-PCR) and first cloned into pGEM-T vector. Then digested by XhoⅠ/HindⅢand identified by PCR.And then subcloned into the retroviral vector pLNCX_2. pLNCX_2-Livin was transferred into packaging cell PT67 after XhoⅠ/HindⅢdigestion and identified by PCR. PT67/pLNCX2-Livin cells were selected. A pure packaging cell line/pLNCX2-Livin cells expressing Livin was cultured. The virus titer was assayed.
Results:816bp Livin cDNA was obtained by RT-PCR amplification. The recombinant vector pLNCX2-Livin containing Livin gene was confirmed. The virus titer of the pure packaging cell line/ PT 67/ pLLNCX2-Livin was 3.9×10~7 IU/ml.
Conclusion:A kind of recombinant retroviral vector pLNCX2-Livin as well as a pure packaging cell line producing high virus titer are constructed successfully.
PART 2 Effects of Livin on Myocardial Ischemiareperfusion Injury in Rats
Objectives: To observe the livin protein expression in rat myocardial after the transfection of retroviral vectors and its effects on ischemia-reperfusion (I/R) injury.
Methods: (1)90 healthy SD rats were divided into Contrl group, I/R group and Livin group. Rats were subjected to 30 min of left coronary artery occlusion followed by 120 min of reperfusion with (Livin group) or without (IR group) treating the rats with retrovirus vector expressing Livin protein by intramyocardial injection 24h before left coronary artery occlusion. Rats in Control group underwent thoracotomy without coronary ligation and injection of vector.
(2) No-reflow area was detected by thioflavin S and the myocardial infarction size was evaluated by TTC dyeing method.
(3)Caspase-3 and Livin expression detected with S? immunohistochemical staining.
(4)Cardiomyocytes apoptosis was determined with terminal deoxynucleotidyl transferase-mediated Dutp-fluorescein nick end labeling( TUNEL) method .
Results:
1.The expression of Livin was detected in myocardial cells by immunohisto- chemistry. The level of Livin was increased by Ischemia-reperfusion injury. There is no statistical significance between IR group (3.10±0.52) and Control group (2.35±0.47).
2.The level of Caspase-3 increased significantly during ischemia-reperfusion; There was significant diference between IR group(103.39±8.24) and Control group (91.39±4.82). The expression of Caspase-3 was decreased after livin injection, there was significant difference between Livin group (97.43±11.15) and IR group.
3. The infarct size (12.80±1.19 vs 20.82±2.82) and the no-reflow area (19.39±6.25 vs 25.93±6.45) were also decreased in Livin groups compared with the IR hearts.
4. Using TUNEL assays, we examined the presence of apoptosis during I/R, which was significantly higher in IR group compared with the control hearts (10.35±3.34 vs 1.1±0.42). Apoptosis rate was significantly reduced after Livin rejection (1.7±1.57) versus IR, but had no difference with control group.
Conclusion
1.There is Livin expression in normal rat myocardial.
2.Retrovirus vector expressing Livin protein could express successfully in rat myocardial.
3.Livin could inhibit caspase-3 expression in myocardial during I/R.
4.Livin inhibits apoptosis, reduces no-reflow area and infartion size in myocardial, and protects the heart against ischemia/reperfusion injury.
PART 3 Caspase-3, Livin mRNA detection by real time polymerase chain reaction
Objectives: To observe the livin and Caspase-3 mRNA expression in rat myocardial with and without retroviral vectors transfection and its effects on ischemia-reperfusion (I/R) injury.
Methods: Both Caspase-3 and Livin mRNA expression were detected by real time polymerase chain reaction (PCR).
Results: The livin mRNA expression was positive in normal rats detected by real time quantification PCR. The livin mRNA expression was (8.41±1.39)×10~(-3) in control group, (7. 45±2. 51)×10~(-3) in vector without livin gene group, (7.82±3.22)×10~(-3) in IR group, (145±89)×10~(-3) in Livin group. Compared with the Control and IR groups, Livin vector transfection significantly increased Livin mRNA expression.The Caspase-3 mRNA expression was (5.12±2.11)×10~(-3) in control group, (6.88±2.31)×10~(-3) in vector without livin gene group. Compared with the control group, I/R significantly increased Caspase-3 mRNA expression [(92.1±34.6)×10~(-3), I/R group , P<0.01], which indicated that reperfusion could increase Caspase-3 mRNA expression. The Caspase-3 mRNA expression was (56.2±21.1)×10~(-3) in Livin group, which was significantly decreased compared to IR group (P<0. 05) . These suggest that Livin overexpression could depress Caspase-3 expression in myocardial during I/R.
Conclusions:
1.There is Livin expression in normal rat myocardial.
2.Retroviral vector expressing Livin protein could express successfully in rat myocardial.
3.Livin could depress caspase-3 expression in myocardial during I/R.
引文
[1] Hanna N, Singh N. Role of caspase in apoptosis and disease. Indian J Physiol Pharmacol 1999,43(2):151-159
[2] Ashhab Y, Alian A, Polliack A, et al. Two splicing variants of a new inhibitor of apoptosis gene with different biological properties and tissue distribution attern. FEBS Lett, 2001, 495(1-2): 56-60.
[3] Lin KM, Lin B, Lianl Y, et al. Combined and individual mitochondrial HSP60 and HSP1O expression in cardiac myocytes protects mitochondrial function and prevents apoptotic cell deaths induced by simulated ischemia reoxygenation. Circulation, 2001,103(13): 1787-1792.
[4] Murtaza I,Wang HX,Feng X,et al. Down regulation of catalase and oxidative modification of protein kinase CK2 lead to the failure of apoptosis repressor with caspase recruitment domain to inhibit cardiomyocyte hypertrophy. J Bioil Chem,2008 Mar 7;283(10):5996-6004.
[5] Gustafsson AB, Gottlieb RA. Heart mitochondria: gates of life and death.Cardiovasc Res, 2008 Feb l;77(2):334-43.
[6] Hariu H, Hirohashi Y, Torigoe T, et al. Aberant expression and potency as a cancer immuotherapy target of inhibitor of apoptosis protein family Livin/ML-IAP in lung cancer.Clin Cancer Res, 2005, 11(3): 1000-1009.
[7] Vucic D,Deshayes K,Ackerly H,et al.SMAC negatively regulates anti-apoptosis activity of melanoma inhibitor of apoptosis(MLIAP).J Biol Chem, 2002, 277:12275-12279.
[8] Ka H, Hunt JS, Temporal and spatial patterns of expression of inhibitors of apoptosis in placentas.Am J Pathol, 2003, 163(2): 413-422
[9] Zhu Z,Wang Y,Ding X,et al. Expression and prognostic significance of Livin in the progression of bladder cancer.J Huazhong Univ Sci Technolog Med Sci, 2008 Feb;28(1):90-2
[10] Gazzaniga P, Gradilone A,Giuliani L, et al. Expression and prognostic significance of Livin, Survivin and other apoptosis related genes in the progression of superficial bladder cancer. Ann Oncol, 2003, 14(1): 85-90
[11]Ashhab Y, Alian A, Polliack A, et al. Two splicing variants of a new inhibitor of apoptosis gene with different biological properties and tissue distribution pattern.FEBS Lett, 2001, 495(1-2): 56-60
[12] Black SC, Huang JQ, Rezaiefar P, et al. Co-localization of the cysteine protease caspase-3 with apoptotic myocytes after in vivo myocardial ischemia and reperfusion in the rat.J Mol Cell cardiol, 1998,30(4) :733-742
[13]Wagener N,Crnkovic-Mertens I,Vetter C,et al.Expression of inhibitor of apoptosis protein Livin in renal cell carcinoma and non-tumorous adult kidney.Br j Cancer,2007 Nov 5;97(9):1271-6.
[14]Enari M, Sakahira H, Yokoyama H, et al. A Caspase-activated DNase that degrades DNA during apoptosis,and its inhibitor ICAD.Nature.1998,391:43-50
[15]Wang H, Tan SS, Wang XY, et al.Silencing livin gene by siRNA leads to apoptosis induction, cell cycle arrest, and proliferation inhibition in malignant melanoma LiBr cells. Acta pharmacol Sin,2007 Dec;28(12):1968-74
[16]Kleinberg L, Lei AK, Florenes VA, et al. Expression of inhibitor-of-apoptosis protein family members in malignant mesothelioma. Hum Pathol, 2007 Jul; 38(7):986-94.
[17]Nachmias B,Lazar I,Elmalech M,et al. Subcellular localization determines the delicate balance between the anti- and pro-apoptotic activity of Livin. Apoptosis,2007 Jul;12(7):1129-42
[18] Chang h,Schimmer AD. Livin/melanoma inhibitor of apoptosis protein as a potential therapeutic target for the treatment of malignancy.Mol Cancer Ther,2007 Jan;6(1):24-30
[19]Vucic D, Stennicke HR, Pisabarro MT, et al. ML-IAP, a novel inhibitor of apoptosis that is preferentially expressed in human melanomas. Curr Biol, 2000,10(21): 1359-1366
[20]Achmias B,Ashhab Y, Bucholtz V, et al. Caspase-ediated cleavage converts Livin from an antiapoptitic to a proapoptotic factor:implications for drug-resistant
??melanoma. Cancer Res, 2003, 63(19): 6340-6349
[21] Gordon GJ, Mani M, Mukhopadhyay 1, et al. Expression patterns of inhibitor ofapoptosis proteins in malignant pleural mesothelioma. J Pathol. 2007,211(4):447-454
[22]Bonlares HA, Zoltoski A J, Yakovlev A, et al. Roles of DNA fragmentation factorand poly(ADP-ribose) polymerase in an amplification phase of tumornecrosis factor infuced apoptosis.J Bio Chemi, 2001,276(41):28185-28192
[23] Wang H,Tan SS,Wang XY,et al. Silencing livin gene by siRNA leads to apoptosisinduction, cell cycle arrest, and proliferation inhibition in malignant melanoma LiBrcells.Acta Pharmacol Sin, 2007 Dec;28(12): 1968-74
[24]Sanna MG, Correia JDS, Ducrey O, et al. IAP Suppression of Apoptosis InvolvesDistinct Mechanisms: the TAK1/JNK1 Signaling Cascade and Caspase Inhibition. MolCell Biol, 2002, 22(6): 1754-1766.
[25]段小军，杨柳，董世武，等．人HIF-1a基因重组逆转录病毒载体构建及鉴定．第 三军医大学学报，2004，26(8)：1639
[1]胡大一，马长生．心脏病学实践2007——新进展与临床案例．北京：人民卫生出版 社．2007：10,19-21．
[2]睢大员，吕忠智，于晓风，等．益心口服液对急性心肌缺血大鼠心梗面积及血液流 变学的影响．中国实验方剂学杂志，1996，2(3)：14-16．
[3]邓汉武．大鼠心肌缺血再灌注损伤实验法．中国药理学通报，1989，5(5)：317-320．
[4]方喜业．医学实验动物学．北京：人民卫生出版社．1995,291-295．
[5] Yaoita H, Ogawa K, Maeham K.et al. Apoptosis in relevent clinicalsituationxontribution of apoptosis in myocardial infraction.Cardiovascular Res, 2000,45:630-641.
[6] Yellon DM, Baxter CF. Protecting the ischemia and reperfused myocardium in actuemyocardial infarction:distant dream or near reality?Heart,2000,83:381-387.
[7] Hearse DJ. Reperfusion of the ischemic myocardium.Journal of Molecular and CellularCardiology,1977,9:605-616.
[8] Fliss H,Gattinger D. Apoptosis in ischemic and reperfused rat myocardium.CirRes,1996,79(5):949-956.
[9] Saraste A,Pulkki K, Kallajok M,et al.Apoptosis in human acute myocardial infar-ction.Circulation,1997,95(2):320-323.
[10]Kasof GM,Gomes BC.Livin,a novel inhibitor of apoptosis protein family member.JBiol Chem,2001,276(5):3238-3246.
[11]Lin JH, Deng G, Huang Q,et al. KIAP,a novel member of the inhibitor of apoptosisprotein family. Biochem Biophys Res Commun, 2000, 279(3):820-831.
[12]Gazzaniga P,Gradilone A,Giuliani L,et al.Expression and prognostic significance ofLivin,Survivin and other apoptosis related genes in the progression of superficialbladder cancer. Ann Oncol, 2003,14(1):85-90.
[13]Ashhab Y, Alian A, Polliack A, et al. Two splicing variants of a new inhibitor ofapoptosis gene with different biologicalproperties and tissue distribution pattern. FEBSLett, 2001,495(1-2):56-60.
[14]Vucic D, Deshayes K, Ackerly H,et al. SMAC negatively regulates the anti- apoptotic activity of melanoma inhibitor of apoptosis(ML-IAP).Biol Chem, 2002,277(14):12275-12279.
[15]Ka H, Hunt JS. Temporal and spatial patterns of expression of inhibitors of apoptosis in placentas.Am J Pathol,2003,163(2):413-422.
[16]Hariu H, Hirohashi Y,Torigoe T,et al. Aberant expression and potency as a cancer immuotherapy target of inhibitor of apoptosis protein familyLivin/ML-IAP in lung cancer.Clin Cancer Res, 2005,ll(3):1000-1009.
[17]邱红,邵淑丽.细胞凋亡的检测方法.高师理科学刊. 2006, 26(1):68-71.
[18]Hanna N, Singh N. Role of caspase in apoptosis and disease.Indian J Physiol Pharmacol 1999,43(2): 151-159.
[19]Higuchi M,Aggarwal BB,Yeh ET,et al. Activation of CPP 32-like protease in tumor necrosis factor-induced apoptosis is dependent on mitochondrial function.Clin Invest,1997,99(7):1751-1758.
[20]Depraetere V, Golstein P. Dismantling in cell death:molecular mechanisms and relationship to caspase activation.Scand J Immunol,1998,47(6):523-531.
[21]Bonlares HA,Zoltoski A J,Yakovlev A,et al. Roles of DNA fragmentation factor and poly(ADP-ribose) polymerase in an amplification phase of tumor necrosis factor infuced apoptosis.J Bio Chemi,2001,276(41):28185-28192.
[22]Idriss HT, Naisnith JH. TNF alpha and TNF receptor super family: structure- function relationships.Microsc ResTech,2000,50(3):184-195.
[23] Black SC, Huang JQ,Rezaiefar P,et al.Co-localization of the cysteine protease caspase-3 with apoptotic myocytes after in vivo myocardial ischemia and reperfusion in the rat. J Mol Cell cardiol,1998,30 (4):733-742.
[24]Yaoita H, Ogawa K, Maehara K, et al. Attenuation of ischemia/reperfusion injury in rats by a caspase inhibitor.Circulation .1998,97:276-281.
[25]Enari M, Sakahira H, Yokoyama H, et al. A Caspase-activated DNase that degrades DNA during apoptosis,and its inhibitor ICAD. Nature. 1998, 391: 43-50.
[26] Mercer EA,Korhonen L,Skoglosa Yet al.NAIP interacts with hippocalcin and protects neurons against calcium-induced cell death through caspase-3-dependent and independent pathways.EMBO J 2000,19(14):3597-3607.
[27]Takahashi R,Deveraux Q,Tamm I,et al. A single BIR domain of XIAP sufficicent for inhibiting caspases.J Biol Chem,1998,273(14):7787-7790.
[28] Sun C,Cai M,Gunasekera AH,et al.NMR structure and mutagenesis of the inhibitor of apoptosis protein XIAP.Nature,1999,401(6755):818-822.
[29]Vucic D,Stennicke HR,Pisabarro MT,et al. ML-IAP,a novel inhibitor of apoptosis that is preferentially expressed in human melanomas. Curr Biol, 2000,10(21): 1359-1366.
[30]Achmias B, Ashhab Y, Bucholtz V, et al. Caspase-ediated cleavage converts Livin from an antiapoptitic to a proapoptotic factor:implications for drug-resistant melanoma.Cancer Res, 2003, 63(19):6340-6349.
[31]Tanaka M,Ito H,Adachi S,et al.Hypoxia induces apoptosis with enhanced expres- sion of Fas antigen messenger RNA in cultured neonatal rat cardiomyocytes. Circ Res,1994,75(3):426-433.
[32] Condorelli G,Roncarati R,Ross J,et al. Heart-targeted overexpression of caspase3 in mice increases infarct size and depresses cardiac function.Proc Natl Acad Sci USA.2001,98(17):9977-82.
[33]Golstein P.Cell Death in us and others.Science.1998; 281:1283.
[34]Kerr JF,Wyllie AH,Currie A R.Apoptosis:a basic biological phenomenon with wide-ranging implications in tissue kinetics.Br J Cancer,1972,26(4):239-257.
[35] Gottlieb RA,Burleson KO,Kloner RA,et al.Reperfusion injury induces apoptosis in rabbit cardiomyocytes.J Clin Invest,1994,94(4):1621-1628.
[36] Olivetti G, Abbi R,Quaini F,et al.Apoptosis in the failing human heart. N Engl J Med,1997,336(16):1131.
[37]Saraste A, Pulkki K, Kallajoki M,et al.Apoptosis in human acute myocardial infarction.Circulation.l997,95(2):320-323.
[38] Cohen GM.Casepase:the executioners of apoptosis.Iochem J 1997,326 (pt1): 1-16.
[39] Zhang KR, Zhang QJ,Li QX,et al.Long-term aerobic exercise protects the heart against ischemia/reperfusion injury via PI3 kinase-dependent and Akt-mediated mechanism.Apoptosis,2007,12(9):1579-1588.
[40]Bhuiyan MS, Fukunaga K.Inhibition of HtrA2/Omi ameliorates heart dysfunction following ischemia/reperfusion injury in rat heart in vivo. Eur J Pharmacol,2007,557(2-3):168-177.
[41]Fu XC, Wang MW,Li SP,et al.Anti-apoptotic effect and the mechanism of orientin on ischemic/reperfused myocardium.J Asian Nat Prod Res,2006,8(3):265-272.
[42]Ruixing Y,AI-Ghazali R,Wenwu L,et al.Pretreatment with producol attenuates cardiomyocyte apoptosis in a rabbit model of ischemia/reperfusion.Scand J ClinLabInvest.2006,66(7):549-558.
[43]Earnshaw WC, Martins LM, Kaufmann SH. Mammalian caspases: structure, activation, substrates,and functions during apoptosis.Annu Rev Biochem, 1999, 68:383-424.
[44]Yang BG, Zander DS, Mehta JL. Hypoxia-reoxygenation-induced apoptosis in cultured adult rat myocytes and the protective effect of platelets and transforming growth factor-β.The Journal of Pharmacology and Experimental Therapertics, 1999,291:733-738.
[45]Jagat N, Pramod P, Eloisa A,et al. Apoptosis in heart failure: Release of Cytc from mitochondrial and activation of spase-3 in human cardiomyopathy.Proc Natl Acad Sci USA,1999,96:8144-8149.
[46] Green DR.Apoptoic pathways:the roads to ruin.Cell,1998,94(6):695-698.
[47]Rumi M, Genzou T, Takuma A, et al. Dynamic process of apoptosis in adult rat cardiomyocytes analyzed using 48-hour videomicroscopy and electron microscopy.Am Pathology,2001,159:683-691.
[48]Cryns V, Yuan J. Proteases to die for. Genes &Dev,1998,12(1):1551-1570.
[49]Song W, Lu XG, Feng QP.Tumolor necrosis factor-ainduces apoptosis via inducible nitric oxide synthase in neonatal mouse cardiomyocytes. Cardiovascular research,2000,45:595-602.
[50]Treena EM, Michelie NG, Chris MC, et al. Endotoxin infusion in rats induces apoptotic and survival pathways in hearts.Am J Physiol Heart Circ Physiol, 2000, 279:2053-2061.
[51]Kloner RA,Ganote CE,Jennings RB.The"no-reflow"phenomenon after temporary coronary occlusion in the dog.J Clin Invest,1974,54:1496-1508.
[52].Schofer J, Monta R, Mathey D.Scintigraphic of the 'no-reflow'phenomenon in human beings after coronary thrombolysis.J Am Coll Cardiol,1985,5:593-598.
[53].Porter TR, Li S, Oster R,et al.The clinical implication of no reflow demonstrated with intravenous perfluorocarbon containing microbubbles following restoration of thrombolysis in myocardial infarction(TIMI)3 flow in patients with acute myocardial infarction. Am J cardiol, 1998, 82:1173-1177.
[54].Ito H, Maruyama A, Iwakura K,et al. Clinical implications of the "no-reflow" phenomenon: a predictor of complications and left ventricular remodeling in reperfused anterior wall myocardial infarction.Circulation, 1996,93:223-228.
[55].Sakuma T,Hayashi Y,sumi K,et al Prediction of short and intermediated prognosis of patients with acute myocardial infarction using myocardial contrast echocardiography one day after recanalization.J Am coll cardial,1998,32:890-897.
[56].Mattichak SJ, Dixon SR, Shannon F,et al.Failed percutaneous coronary intervention:A decade of experience in 21,000 patients. Catheter Cardiovasc interv,2008 Jan 29;71(2):131-137
[57].Movahed MR, Butman SM. The pathogenesis and treatment of no-reflow occurring during percutaneous coronary intervention.Cardiovasc Revasc med,2008 Jan-Mar;9(1):56-61
[58].Timurkaynak T,Arslan U,Balcioglu S,et al. Long term tirofiban infusion before percutaneous coronary intervention in patients with angiographically massive intracoronary thrombus.Saudi Med J, 2008 Jan;29(1):42-7.
[59].Patel S,Hermiller J. Embolic protection: the FilterWire EZtrade mark Embolic Protection System.Expert Rev med Devices,2008 Jan;5(1):19-24
[60].Daemen M A,de Vries B,van'tVeer C,et al. Apoptosis and chemokine induction after renal ischemia reperfusion. Transplantation, 2001,71(7): 1007.
[61].Matsuo H,Watanabe S,Watanabe T,et al Prevention of no-reflow/slow-flow phenomenon during rotational atherectomy--a prospective randomized study comparing intracoronary continuous infusion of verapamil and nicorandil.Am Heart J,2007 Nov;154(5):994.e1-6.
[62].Scarabelli T,Stephanou A,Rayment N,et al. Apoptosis of endothelial cells precedes myocyte cell apoptosis in ischemia/reperfusion injury.Circulation, 2001, 104(3):
??253-256．
[63]．魏宇淼，洪梅，曾秋棠，等．局部冠状动脉循环内皮细胞凋亡及高凝状态在急性 心肌梗死直接经皮冠状动脉腔内成形术再灌注治疗无复流中的作用。临床心血管 病杂志，2006，22(5)：259
[1]曹林生．心肌缺血再灌注损伤．毛焕元，曹林生主编．心脏病学．第二版．北京：人民 卫生出版社．2005：10，1035
[2] Iliodromitis EK,Aazou L,Kremastinos DT. Ischemic preconditioning: protectionagainst myocardial necrosis and apoptosis.Vasc Health Risk Manag,2007;3(5):629-37.
[3] Hearse DJ. Reperfusion of the ischemic myocardium.Journal of Molecular and CellularCardiology,1977,9:605-616
[4] Hu C,ChenJ,Dandapat A,et al. LOX-1 abrogation reduces myocardialischemia-reperfusion injury in mice.J Mol Cell Cardiol. 2008 Jan; 44(1):76-83.
[5] Khan M,Mohan VK,Kutala VK,et al.Cardioprotection by sulfaphenazole, acytochrome p450 inhibitor: mitigation of ischemia-reperfusion injury by scavenging ofreactive oxygen species.J Pharmacol Exp Ther. 2007 Dec;323 (3):813-21.
[6] Gustafsson AB, Gottlieb RA. Heart mitochondria: gates of life and death.CardiovascRes, 2008 Feb l;77(2):334-43.
[7] Saraste A, Pulkki K, Kallajok M, et al. Apoptosis in human acute myocardialinfarction.Circulation,1997, 95(2): 320-323
[8] Kasof GM, Gomes BC. Livin, a novel inhibitor of apoptosis protein familymember. J Biol Chem, 2001, 276(5): 3238-3246
[9]李国营，刘靖，田素民，等．兔实验性心肌缺血再灌注损伤与心肌细胞凋亡的 关系[J]．解剖学研究，2004，26(1)：5．
[10]邓汉武．大鼠心肌缺血再灌注损伤实验法．中国药理学通报．1989，5：317-320
[11]方喜业． 医学实验动物学．第一版．北京：人民卫生出版社．1995，291-295
[12] Kerr JF,Wyllie AH,Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics.Br J Cancer, 1972, 26(4): 239-257
[13]陈主初．病理生理学．北京：人民卫生出版社，2005：26
[14] Fliss H. Accelerated apoptosis in reperfused myocardium: friend of foe.Basic Res Cardial, 1998,93(2):90.
[15] Hanna N, Singh N, Role of caspase in apoptosis and disease. Indian J Physiol Pharmacol 1999,43 (2):151-159
[16] Stanic I.Ccetrullo S,Facchini A,et al.Effect of the polyamine analogue N(1),N(11)-diethylnorspermine on cell survival and susceptibility to apoptosis of human chondrocytes.J Cell Physiol,2008 Jan 216 (1) : 153-61
[17] Wang H, Tan SS, Wang XY, et al.Silencing livin gene by siRNA leads to apoptosis induction, cell cycle arrest, and proliferation inhibition in malignant melanoma LiBr cells. Acta pharmacol Sin,2007 Dec;28(12):1968-74
[18] Wei Song, Xiangru Lu, Qingping Feng. Tumolor necrosis factor-a induces apoptosis via inducible nitric oxide synthase in neonatal mouse cardiomyocytes.Cardiovascular research, 2000,45:595-602
[19] Treena EM, Michelie NG, Chris MC, et al. Endotoxin infusion in rats induces apoptotic and survival pathways in hearts. Am J Physiol Heart Circ Physiol, 2000, 279:2053-2061
[20] Zhang KR, Zhang QJ,Li QX, et al.Long-term aerobic exercise protects the heart against ischemia/reperfusion injury via PI3 kinase-dependent and Akt-mediated mechanism. Apoptosis,2007,12(9):1579-1588
[21] Bhuiyan MS , Fukunaga K.Inhibition of HtrA2/Omi ameliorates heart dysfunction following ischemia/reperfusion injury in rat heart in vivo.Eur J Pharmacol, 2007, 557:168-177
[22] Fu XC, Wang MW, Li SP,et al. Anti-apoptotic effect and the mechanism of orientin on ischemic/reperfused myocardium. J Asian Nat Prod Res. 2006; 8(3):265-272
[23] Ruixing Y, AI-Ghazali R, Wenwu L,et al. Pretreatment with producol attenuates cardiomyocyte apoptosis in a rabbit model of ischemia reperfusion. Scand J Clin Lab Invest. 2006;66(7):549-558
[24] Kleinberg L, Lei AK, Florenes VA, et al. Expression of inhibitor-of-apoptosis protein family members in malignant mesothelioma. Hum Pathol, 2007 Jul; 38(7):986-94.
[25] Mercer EA, Korhonen L, Skoglosa Y, et al. NAIP interacts with hippocalcin and protects neurons against calcium-induced cell death through caspase-3-dependent and independent.Pathways. EMBO J 2000, 19(14): 3597-3607
[26] nachmias B,Lazar I,Elmalech M,et al. Subcellular localization determines the delicate balance between the anti- and pro-apoptotic activity of Livin. Apoptosis,2007 Jul;12(7):1129-42
[27] Chang H,Schimmer AD. Livin/melanoma inhibitor of apoptosis protein as a potential therapeutic target for the treatment of malignancy.Mol Cancer Ther,2007 Jan;6(1):24-30
[28]VucicD, Stennicke HR, Pisabarro MT, etal. ML-IAP, a novel inhibitor of apoptosis that is preferentially expressed in human melanomas. Curr Biol, 2000,10(21): 1359-1366
[29] Achmias B,Ashhab Y, Bucholtz V, et al. Caspase-ediated cleavage converts Livin from an antiapoptitic to a proapoptotic factor: implications for drug-resistant melanoma. Cancer Res, 2003, 63(19): 6340-6349
[30] Gordon GJ, Mani M, Mukhopadhyay 1, et al. Expression patterns of inhibitor of apoptosis proteins in malignant pleural mesothelioma. J Pathol. 2007,211(4):447-454
[31] Bonlares HA, Zoltoski A J, Yakovlev A, et al. Roles of DNA fragmentation factor and poly(ADP-ribose) polymerase in an amplification phase of tumor necrosis factor infuced apoptosis.J Bio Chemi, 2001,276(41):28185-28192
[32] Idriss HT, Naisnith JH, TNF alpha and TNF receptor super family: structure-function relationships. Microsc Res Tech, 2000,50(3): 184-195
[33] Yan H,Brouha B,Liu T,et al. Proteolytic cleavage of Livin (ML-IAP) in apoptotic melanoma cells potentially mediated by a non-canonical caspaseJ Dermatol Sci,2006 Sep;43(3):189-200.
[34] Sanna MG, Correia JDS, Ducrey O, et al. IAP Suppression of Apoptosis Involves Distinct Mechanisms: the TAK1/JNK1 Signaling Cascade and Caspase Inhibition. Mol Cell Biol, 2002, 22(6): 1754-1766.
[35] Gottlieb RA, Burleson KO, Kloner RA, et al. Reperfusion injury induces apoptosis in rabbit cardiomyocytes. J Clin Invest, 1994, 94(4): 1621-1628
[36] Olivetti G, Abbi R, Quaini F, et al. Apoptosis in the failing human heart. N Engl J Med, 1997,336(16):1131
[1] Iliodromitis EK,Aazou L,Kremastinos DT. Ischemic preconditioning: protection against myocardial necrosis and apoptosis.Vasc Health Risk Manag,2007;3(5):629-37.
[2] Hu C,ChenJ,Dandapat A,et al. LOX-1 abrogation reduces myocardial ischemia-reperfusion injury in mice.J Mol Cell Cardiol. 2008 Jan;44(1):76-83.
[3] Matsuo H,Watanabe S,Watanabe T,et al Prevention of no-reflow/slow-flow phenomenon during rotational atherectomy--a prospective randomized study comparing intracoronary continuous infusion of verapamil and nicorandil.Am Heart J,2007 Nov;154(5):994.el-6.
[4] Kamoda y,Fujino Y,Tanioka y,et al. Ischemically damaged heart after preservation by the cavitary two-layer method as a possible donor in rat heart transplantation. J Heart Lung Transplant,2007 Dec;26(12):1320-5.
[5] Hearse DJ. Reperfusion of the ischemic myocardium[J].Journal of Molecular and Cellular Cardiology,1977,9:605-616
[6] Khan M,Mohan VK,Kutala VK,et al.Cardioprotection by sulfaphenazole, a cytochrome p450 inhibitor: mitigation of ischemia-reperfusion injury by scavenging of reactive oxygen species.J Pharmacol Exp Ther. 2007 Dec;323(3):813-21.
[7] Gustafsson AB,Gottlieb RA. Heart mitochondria: gates of life and death.Cardiovasc Res,2008 Feb 1;77(2):334-43.
[8] Mani K. Programmed cell death in cardiac myocytes: strategies to maximize post-ischemic salvage.Heart Fail Rev,2008 Jan 4: 193
[9] Saraste A, Pulkki K, Kallajok M, et al. Apoptosis in human acute myocardial infarction.Circulation,1997, 95(2): 320-323
[10]Kerr JF,Wyllie AH,Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer, 1972,26(4):239-257
[11]Penna C,mancardi D,Raimondo S,et al. The paradigm of postconditioning to protect the heart.J Cell Mol med, 2007 Dec 20
[12]Woller G,Brandt E,Mittelstadt J.et al. Platelet factor 4/CXCL4-stimulated human monocytes induce apoptosis in endothelial cells by the release of oxygen radicals. J leukoc Biol, 2008 Jan 18.
[13]Juhasz B, Thirunavukkarasu M, pant R,et al.Bromelain induces cardioprotection against ischemia reperfusion injury through Akt/Foxo pathway in rat myocardium. Am J PhysiolHeart Circ Physiol, 2008 Jan 11.
[14]Fliss H. Accelerated apoptosis in reperfused myocardium: friend of foe.Basic Res Cardial, 1998,93(2):90.
[15] 15. Wu S, Xing W,Peng J,et al.Tumor transfected with CCL21 enhanced reactivity and apoptosis resistance of human monocyte-derived dendritic cells. Immunobiology, 2008,213(5): 417-26
[16]Nizamutdinova IT,Jin YC,Kim JS,et al.Paeonol and Paeoniflorin, the Main Active Principles of Paeonia albiflora, Protect the Heart from Myocardial Ischemia/Reperfusion Injury in Rats. Planta Med.2008 Jan;74(1):14-8.
[17]Cosentino F,francia PCamici GG,et al. Final Common Molecular Pathways of Aging and Cardiovascular Disease. Role of the p66Shc Protein. Arterioscler Thromb Vasc Biol,2007 Dec 27
[18] Rossi MA, Souza AC. Is apoptosis a mechanism of cell death of cardiomyocytes in chronic chagasic myocarditis? Cardiology, 1999, 68(3): 325-331
[19]Dumont EA, Reutelingsperger CP, Smits JF, et al. Real-time imaging of apoptotic cell-membrane changes at the single-cell level in the beating murine heart. Nat-Med,2001, 7(12): 1352-1355
[20]Umansky SR, Tomei LD. Apoptosis in the heart. Adv Pharmacol, 1997,41:383-407
[21]Haunstetter A, Izumo S. Apoptosis: basic mechanisms and implications for cardiovascular disease.Circ Res,1998,82(11):1111-1129
[22]Kajstura J, Cheng W, Reiss K, et al. Apoptotic and necrotic myocyte cell deaths are independent contributing variables of infarct size in rats. Lab Invest, 1996, 74(1):86-107
[23]Tanaka M, Ito H, Adachi S, et al. Hypoxia induces apoptosis with enhanced expression of Fas antigen messenger RNA in cultured neonatal rat cardiomyocytes.
??Circ Res, 1994, 75(3): 426-433
[24] Fliss H, Gattinger D. Apoptosis in ischemic and reperfused rat myocardium. Cir Res,1996, 79(5): 949-956
[25] Olivetti G, Abbi R, Quaini F, et al. Apoptosis in the failing human heart.N Engl JMed, 1997,336(16): 1131
[26]Scarabelli T, Stephanou A, Rayment N, et al. Apoptosis of endothelial cells precedesmyocyte cell apoptosis in ischemia/ reperfusion injury. Circulation, 2001, 104(3):253-256
[27] Cheng W, Li B, Kajstura J, et al. Stretch-induced programmed myocyte cell death. JClin Invest, 1995,96,2247-2259
[28]Khanna N, Singh N, Role of caspase in apoptosis and disease. Indian J PhysiolPharmacol 1999,43 (2) :151-159
[29]Cohen GM.Casepase: the executioners of apoptosis Biochem J 1997, 326(pt1):1-16
[30]Higuchi M, Aggarwal BB, Yeh ET, et al. Activation of CPP 32-like protease intumor necrosis factor-induced apoptosis is dependent on mitochondrial function.JClin Invest, 1997, 99 (7) :1751-1758
[31]Depraetere V, Golstein P. Dismantling in cell death: molecular mechanisms andrelationship to caspase activation. Scand J Immunol, 1998,47(6):523-531
[32]Bonlares HA, Zoltoski A J, Yakovlev A, et al. Roles of DNA fragmentation factor andpoly(ADP-ribose)polymerase in an amplification phase of tumor necrosis factorinfuced apoptosis. J Bio Chemi, 2001,276(41):28185-28192
[33]吕 建， 陈智超。Livin与肿瘤研究新进展．中华肿瘤防治杂 志，2006，13(17)：1347-1350．
[34] Nunez G, Benedict MA , HU Y, et al. Caspases: the proteases of the apoptoticpathway. Oncogene, 1998, 17 (25) :3 237-3 245
[35]Tschopp J,Martinon F,Burns K: NALPs: a novel protein family involved ininflammation. Nat Rev Mol Cell Biol 2003,4: 95-104.
[36]Mizoguchi K, Maeta H, Yamamoto A, et al. Amelioration of myocardial globalischemia/reperfusion injury with volume-regulatory chloride channel inhibitors in vivo . J.Transplantation, 2002, 73(8): 1185-1193.
[37]Lin KM, Lin B, Lianl Y, el al. Combined and individual mitochondrial HSP60 and HSP10 expression in cardiac myocytes protects mitochondrial function and prevents apoptotic cell deaths induced by simulated ischemia reoxygenation. Circulation, 2001,103(13): 1787-1792.
[38] Nunez G, Benedict M A, Hu Y,et al.Caspases:the protease of the apoptotic pathway.Oncogene,1998,17(25):3237-3247.
[39]Reed JC, Tomaselli KJ. Drug discovery opportunities from apoptosis research. Curr Opin Biotechnol, 2000,11(6): 586-592
[40]Boatright KM, Renatus M, Scott FL, et al. A unified model for apical caspase activation. Mol Cell 2003 ,11:529 - 541
[41] Green DR, Reed JC. Mitochondria and apoptosis. Science. 1998, 281 (5381) :1309-1312
[42]Liu X, Zou H, Slaughter C, et al. DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. cell, 1997,89(2): 175-184
[43]Enari M, Sakahira H, Yokoyama H, et al. A caspase-activated DNase that degrades DNA during apoptosis and its inhibitor ICAD. Nature, 1998, 391: 43-50
[44]Idriss HT, Naisnith JH, TNF alpha and TNF receptor super family: structure-function relationships. Microsc Res Tech, 2000, 50(3): 184-195
[45] Black SC, Huang JQ, Rezaiefar P, et al. Co-localization of the cysteine protease caspase-3 with apoptotic myocytes after in vivo myocardial ischemia and reperfusion in the rat. J Mol Cell cardiol, 1998, 30 (4) :733-742
[46]Rideout HJ, Stefanis L. Caspase inhibition: a potential therapeutic strategy in neurological diseases. Histol Histopathol, 2001,16 (3) :895-908
[47]Wang J, Zhen L, Klug M G, et al. Incolvement of caspase-3 and 8-like protenses in ceramide-induced apoptosis of cardiomyocytes. J Card Fail ,2000, 6 (3) :243-249
[48] Holly TA, Drincic A, Byun Y, et al. Caspase inhibition reduces myocyte cell death induced by myocardial ischemia and reperfusion in vivo. J Mol Cell Cardiol ,1999, 31 (9): 1709-1715
[49]Condorelli G, Roncarati R,Ross J, et al. Heart-targeted overexpression of caspase-3 in mice increases infarct size and depresses cardiac function. Proc Natl Acad Sci USA. 2001, 98 (17) :9977-82
[50] Zhang KR, Zhang QJ,Li QX, et al.Long-term aerobic exercise protects the heart against ischemia/reperfusion injury via PI3 kinase-dependent and Akt-mediated mechanism.Apoptosis, 2007,12(9):1579-1588
[51]Bhuiyan MS , Fukunaga KJnhibition of HtrA2/Omi ameliorates heart dysfunction following ischemia/reperfusion injury in rat heart in vivo.Eur J Pharmacol,2007,557(2-3): 168-177
[52]Fu XC, Wang MW, Li SP,et al. Anti-apoptotic effect and the mechanism of orientin on ischemic/reperfused myocardium. J Asian Nat Prod Res. 2006; 8(3):265-272
[53]Ruixing Y, AI-Ghazali R, Wenwu L,et al. Pretreatment with producol attenuates cardiomyocyte apoptosis in a rabbit model of ischemia/reperfusion. Scand J Clin Lab Invest. 2006; 66(7): 549-558.
[54]Lin JH, Deng G, Huang Q, et al. KIAP, a novel member of the inhibitor of apoptosis protein family. Biochem Biophys Res Commun, 2000, 279(3): 820-831
[55] Vucic D, Stennicke HR, Pisabarro MT, et al. ML-IAP, a novel inhibitor of apoptosis that is preferentially expressed in human melanomas. Curr Biol, 2000, 10(21):1359-1366
[56]Schofer J, Monta R,Mathey D.Scintigraphic of the 'no-reflow'phenomenon in human beings after coronary thrombolysis.J Am Coll Cardiol,1985,5:593-598.
[57]Ashhab Y, Alian A, Polliack A, et al. Two splicing variants of a new inhibitor of apoptosis gene with different biological properties and tissue distribution pattern. FEBS Lett, 2001, 495(1-2): 56-60.
[58]Hauser HP, Bardroff M, Pyrowolaksis G, etal. A giant ubiquitin-conjugating engyme related to LAP apoptosis. J Cell Biol, 1998, 141(6): 1415-1422
[59]Gazzaniga P, Gradilone A, Giuliani L, et al. Expression and prognostic significance of Livin, Survivin and other apoptosis related genes in the progression of superficial bladder cancer. Ann Oncol, 2003, 14(1): 85-90
[60] Crnkovic-Mertens I, Hoppe-Seyler F, Butz K. Induction of apoptosis in tumor cells by siRNA-mediated silencing of the Livin/ML-IAP/KIAP gene. Oncogene, 2003,22(51): 8330-8336
[61]Ashhab Y, Alian A, PolIiack A, et al. Two splicing variants of a new inhibitor of apoptosis gene with different biological properties and tissue distribution pattern. FEBS Lett, 2001, 495(1-2): 56-60
[62]Kasof GM, Gomes BC. Livin, a novel inhibitor of apoptosis protein family member.J Biol Chem, 2001, 276(5): 3238-3246
[63]Hariu H, Hirohashi Y,Torigoe T,et al .Aberrant expression and potency as a cancer immunotherapy target of inhibitor of apoptosis protein family. Livin/ML-IAP in lung cancer.Clin Cancer Res,2005,ll:1000-1009.
[64]Vucic D, Deshayes K,Ackerly H,et al.SMAC negatively regulates anti-apoptosis activity of melanoma inhibitor of apoptosis(MLIAP).J Biol Chem, 2002, 277:12275-12279.
[65] Ka H, Hunt JS. Temporal and spatial patterns of expression of inhibitors of apoptosis in human placentas.Am J Pathol,2003,163:413-422.
[66] Gordon GJ, Mani M, Mukhopadhyay 1, et al. Expression patterns of inhibitor of apoptosis proteins in malignant pleural mesothelioma. J Pathol. 2007,211(4):447-454
[67]Green DR.Apoptoic pathways: the roads to ruin. Cell, 1998,94 (6):695-698
[68]Cryns V,Yuan J. Proteases to die for. Genes & Dev, 1998, 12(l):1551-1570
[69]Sanna MG, da Silva Correia J, Ducrey O et al. IAP suppression of apoptosis involves distinct medhanisms:The TAK1/JNK1 Signaling cascade and caspase inhibition. J Mol Cell Biol, 2002, 22(6): 1754-1766
[70]Stehlik C, de Martin R, Kumabashiri T, et al. Nuclear factor(NF)-kappa B-regulated X-chromosome-linked IAP gene expression protects endothelial cells from tumor necrosis factor alpha-induced apoptosis. J Exp Med, 1998,188(1): 211-216
[71]Rideout HJ, Stefanis L. Caspase inhibition: a potential therapeutic strategy in neurological diseases. Histol Histopathol , 2001,16 (3) :895-908
[72]Wang J , Zhen L, Klug M G, et al . Incolvement of caspase-3 and 8-like pro tenses in ceramide-induced apoptosis of cardiomyocytes. J Card Fail ,2000, 6 (3) :243-249
[73]Wagener N,Crnkovic-Mertens I,Vetter C,et al.Expression of inhibitor of apoptosis protein Livin in renal cell carcinoma and non-tumorous adult kidney.Br J Cancer,2007 Nov5;97(9):1271-6.
[74]Boaz N,Yaqoub A,Vered B,et al.Caspase-mediated cleavage converts livin from an antiapoptotic to proapoptotic factor: implications for drug-resistant melanoma.2003,63(10): 6340.
[75] Anderson MH, Reker S, Becker JC, et al. The Melanoma inhibitor of apoptosis protein: a target for spontaneous cytotoxic T cell respontaneous. J of Investigative Dermatology, 2004,122: 392-399
[76]Wang H,Tan SS,Wang XY, et al.Silencing livin gene by siRNA leads to apoptosis induction, cell cycle arrest, and proliferation inhibition in malignant melanoma LiBr cells. Acta pharmacol Sin,2007 Dec;28(12):1968-74
[77] Chang h,Schimmer AD. Livin/melanoma inhibitor of apoptosis protein as a potential therapeutic target for the treatment of malignancy.Mol Cancer Ther,2007 Jan;6(1):24-30