颅咽管瘤超微结构观察及Caspase-3，-9基因的表达和意义
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摘要
目的 探讨颅咽管瘤超微结构及Caspase-3，-9在颅咽管瘤中的表达和意义。
方法 1 采用Tunel法检测颅咽管瘤及正常脑组织的细胞凋亡情况，同时取4例新鲜颅咽管瘤组织行透射电镜观测超微结构及细胞凋亡。2 采用免疫组化SP法检测50例颅咽管瘤及10例正常对照脑组织中Caspase-3，-9的表达水平，按1996年《全国免疫组织化学技术与诊断标准化专题研讨会》标准对结果判定，并进行统计分析。
结果 1 HE观察：(1)颅咽管瘤釉质上皮型织学特点：组织内上皮成分呈条索状或巢状，上皮巢的周边细胞呈栅栏状排列，为排列整齐的柱状或矮柱状基底细胞，巢内鳞状细胞和星状细胞；(2)鳞形上皮型：实性组织由成片分化好的鳞状上皮组成，肿瘤细胞与间质细胞构成片状、乳头状或分枝状小梁，有些类似皮肤的乳头瘤或基底细胞乳头瘤。2 电镜观察：电镜下细胞核内核仁明显，染色质分布均匀，核膜不规则，细胞浆内线粒体、粗面内质网多，细胞膜有不规则少量突起。釉质上皮型瘤胞浆内中核糖体丰富，基底膜增厚，其上附着丰富的糖元。鳞形上皮型瘤细胞内联结体及瘤细胞间的桥粒均明显多于釉质上皮型，桥粒将表皮型上皮细胞互相紧密联结。瘤细胞坏死相对较多。仅可见1个典型凋亡细胞，其胞核呈椭圆形，细胞核内染色质边集，核膜及细胞膜完整。3 Tunel观察颅咽管瘤及正常脑组织细胞凋亡：结果显示颅咽管瘤釉质上皮型、
Objective: To investigate the ultrastructure of craniopharyngioma and the exprssion and significance of Caspase-3,-9 in craniopharyngiomas
Methods: 1) The cell apoptosis in craniopharyngioma and normal brain tissue was inspected by mean of TUNEL technique and four cases of fresh craniopharyngioma samples was used to examine by means of electromicroscope. 2) The expression of Caspase-3, -9 were detected in 50 cases of craniopharyngiomas and 10 cases of normal brain tissues by immunohistochemical SP technique and the outcome were determined by the standard of China Pathology Magazine Editorial Committee about The Special Seminor of Nationwide Iramuohistochemistry Technique and Diagnosis Standardization instituted in 1996 and were analyzed by statistical method.
Results: 1 HE observation: (1) Histological characteristics of adamantine epithelial craniopharyngiomas: the epithelial components looked like strips or nests .Basis cells looked like Column or short column and arranged in order like barriers in the circumjacent region . Squamous and star cells were located inside the nest. (2)Histological characteristics of squamous papillary craniopharyngiomas: solid parts consisted of sheet squamous
方法 1 采用Tunel法检测颅咽管瘤及正常脑组织的细胞凋亡情况，同时取4例新鲜颅咽管瘤组织行透射电镜观测超微结构及细胞凋亡。2 采用免疫组化SP法检测50例颅咽管瘤及10例正常对照脑组织中Caspase-3，-9的表达水平，按1996年《全国免疫组织化学技术与诊断标准化专题研讨会》标准对结果判定，并进行统计分析。
结果 1 HE观察：(1)颅咽管瘤釉质上皮型织学特点：组织内上皮成分呈条索状或巢状，上皮巢的周边细胞呈栅栏状排列，为排列整齐的柱状或矮柱状基底细胞，巢内鳞状细胞和星状细胞；(2)鳞形上皮型：实性组织由成片分化好的鳞状上皮组成，肿瘤细胞与间质细胞构成片状、乳头状或分枝状小梁，有些类似皮肤的乳头瘤或基底细胞乳头瘤。2 电镜观察：电镜下细胞核内核仁明显，染色质分布均匀，核膜不规则，细胞浆内线粒体、粗面内质网多，细胞膜有不规则少量突起。釉质上皮型瘤胞浆内中核糖体丰富，基底膜增厚，其上附着丰富的糖元。鳞形上皮型瘤细胞内联结体及瘤细胞间的桥粒均明显多于釉质上皮型，桥粒将表皮型上皮细胞互相紧密联结。瘤细胞坏死相对较多。仅可见1个典型凋亡细胞，其胞核呈椭圆形，细胞核内染色质边集，核膜及细胞膜完整。3 Tunel观察颅咽管瘤及正常脑组织细胞凋亡：结果显示颅咽管瘤釉质上皮型、
Objective: To investigate the ultrastructure of craniopharyngioma and the exprssion and significance of Caspase-3,-9 in craniopharyngiomas
Methods: 1) The cell apoptosis in craniopharyngioma and normal brain tissue was inspected by mean of TUNEL technique and four cases of fresh craniopharyngioma samples was used to examine by means of electromicroscope. 2) The expression of Caspase-3, -9 were detected in 50 cases of craniopharyngiomas and 10 cases of normal brain tissues by immunohistochemical SP technique and the outcome were determined by the standard of China Pathology Magazine Editorial Committee about The Special Seminor of Nationwide Iramuohistochemistry Technique and Diagnosis Standardization instituted in 1996 and were analyzed by statistical method.
Results: 1 HE observation: (1) Histological characteristics of adamantine epithelial craniopharyngiomas: the epithelial components looked like strips or nests .Basis cells looked like Column or short column and arranged in order like barriers in the circumjacent region . Squamous and star cells were located inside the nest. (2)Histological characteristics of squamous papillary craniopharyngiomas: solid parts consisted of sheet squamous
引文
1 李晓强、漆松涛等．颅咽管瘤中Bcl-2蛋白的表达及意义．中国神经精神疾病杂志，2004，30(1)：50—52．
2 Reed JC. Dysregulation of apoptosis in cancer[J].J Chin Oncol, 1999, 17: 2941—2953。
3 Hengarter MO. The biochemistry of apoptosis[J]. Nature, 2000, 407: 770-776.
4 Yang J, Liu XS, Bhalla K, et al. Science, 1997; 275: 1129—1132.
5 Adams JM, Copy S. The Bcl-2 protein family: arbiters of cell survival[J]. Science, 1998, 281: 1332—1336.
6 Wright ME, Han DK, Hockenbery DM. Caspase-3 and inhibitor of apoptosis protein interaction in saccharomyces cerevisiae and mammalian cells. FEBS Lett, 2000, 481: 13—14.
7 Deveraux Q. L., Reed J. C. IAP family proteins-suppressors of apoptosis [J]. Gene Develp, 1999; 13 (3): 239—252.
8 Raff M. Cell suicide for beginners[J]. Nature, 1998, 396(6707): 119—122.
9 Fridlander RM, Yuan J.ICE, neuronal apoptosis and neurodegeneration[J]. Cell Death Differ, 1998, 5(10): 823—831.
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12 Askenazi A, Dixit VM, Death receptors: signaling and modulation[J]. Science, 1998, 281: 1305—1308.
13 Hu S, Snipas SJ, Vincenz C, et al. Caspase-14 is a novel developmentally regulated protease[J]. J Biol Chem, 1998, 273: 29648—29653.
14 Hirata H, Takahashi A, Kobayashi S, et al. Caspases are activated in a branched protease cascade and control distinct downstream processes in Fas-induced apoptosis[J]. J Exp Med, 1998, 187(4): 587—600.
15 Stennicke HR, Salvesen GS. Properties of the caspase[J]. Bichim Biophys Acta, 1998, 1387: 17—31.
16 中华病理学杂志编辑委员会，《全国免疫组织化学技术与诊断标准化专题研讨会》会议记要[J]．中华病理学杂志，1996；25(6)：326．
17 徐建国．颅咽管瘤的动物模型的建立及肿瘤生物学特性的研究．四川大学临床医学博士专业学位论文．B202658．2005，4．
18 Schut, Louis MD, Discussant, et al. Management of a Pediatric Hypothalamic Mass. [Miscellaneous Article]. Neurosurgery. 1996, Apr, 38(4): 806-811.
19 Liu JM, Garonzik IM, Eberhart CG, et al. Ectopic recurrence of craniopharygioma after an interhemispheric transcallosal approacha: case report. [Revew]. Neurosurgery. 2002 Mar, 50(3): 639-644; discussion644-645.
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21 Krajewaka M, Wang HG; Krajewaki S, et al. Immunohistochemistry of in vivo patterns of expression of CPP32(caspase3),a cell death protease. Cancer Res, 1997, 57: 1605—1613.
22 孙宝华，张俊，王保菊等．Caspase3基因表达与原发性肝细胞癌凋亡 无关．中华肿瘤杂志．2000，22(1)39—41．
23 文亚渊，刘宝华．基因Survivin、Caspase-3在胃癌发生中的作用．消化外科，2004，3(6)：415—418．
24 文亚渊，刘宝华，付涛．基因Survivin、Caspase-9在胃癌组织中的表达及与胃癌发生的关系．中国肿瘤临床，2005，32(17)：969—972．
25 高红，王常林，蔡炜嵩等．Survivin、Caspase-3mRNA在肾母细胞瘤中的表达及意义．中国当代儿科杂志，2004，6(5)：381—384．
26 周舟，王小华，朱光旭等．Caspase-3、9表达上调参与缺氧诱导心肌细胞凋亡．第三军医大学学报，2005，27(3)：185-188．
27 徐强，司良毅，赵小兰等．大鼠心肌缺血再灌注时心肌细胞凋亡caspase-3活性变化规律及药物对其影响．第三军医大学学报，2003，25(15)：1335-1357．
28 易成腊，陈安民，徐卫国等．大鼠脊髓损伤后Caspase-3表达和神经细胞凋亡相关性研究．创伤外科杂志．2004，6(3)：171—174．
29 Springer JE, Azbil RD, Knapp PE. Activation of the Caspase-3 apoptotic cascade in traumatic spinal cord injury[J]. Nat Med, 1999, 5(8): 943—946.
30 Cheng Y, Deshmukh M, Dcosta A, et al. Caspase inhibitor affords neuroprotection with delayed administration in a rat model of neonatal hypoxicischemic brain injury. J clin invest, 1998, 101(9): 1992.
31 郭燕舞，杨鹏飞，徐如祥．Caspase-3在脑损伤时的作用．广东医学．2004，25(9)：1026—1028．
32 James J, Velier A, Ellison K, et al. Caspase-8 and Caspase-3 Are Expressed by Different Populations of Cortical Neurons Undergoing Delayed Cell Death after Focal Stroke in the Rat. The Journal of Neuroscience, July 15, 1999, 19(14): 5932—5941.
33 郭云良，陈红兵，高英茂．大鼠脑缺血再灌注后神经细胞凋亡与Caspase-3和Caspase-9基因表达．解剖学杂志．2003，26(5)：430—434．
34 Gong C, Boulis N, Qian J, et al. Intracerebral hemorrhage-induced neuronal death. Neurosurgy. 2001, 48 (4): 875.
35 吴家幂，李珺．Caspase-3在脑出血后神经细胞凋亡中的作用．放射免疫学杂志．2005，18(2)：135—137．
36 Emery E, Aldana P, Bunge MB, et al. Apoptosis after traumatic human spinal cord injury[J]. J Neurosurg, 1998, 89(6): 911—920.
37 王文昭，戚基萍，王得升等．人脑出血后海马神经元损伤与Caspase-3的关系．中风与神经疾病杂志．05．22(2)：107—110．
38 Hoys, TsaiPW, Yu C. F, et al. Ketoconazole-induced apoptosis through p53-dependent pathway in human colorectal and hepatocelluarcarcinoma cell line[J]. Toxicol-APP1-pharmacol, 1998, 153: 39—47.
39 杨建青，黄柏英，吕新生．Caspase-9在氟尿嘧啶诱导的肝癌细胞凋亡中的作用．中国普通外科杂志．2003，12(3)：173—175．
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41 耿伟，沈志强，陈水清等．凋亡相关基因Caspase-9，Bax在大肠腺癌和大肠癌的表达及意义．胃肠病学和肝病学杂志．，2004，13(6)：635—638．
42 Kondo S,Bama Bp,Morimura T,et al.Interleukin-1 B coverting enzyme mediates cisplatin-induced apoptosis in malignant glioma cells [J]. Cancer Res,1995,55:6166—6171.
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1. Reed JC.Dysregulation of apoptosis in cancer[J].J Chin Oncol, 1999, 17: 2941—2953.
2. Hengarter MO. The biochemistry of apoptosis [J]. Nature,2000,407:770-776.
3. Yang J, Liu XS, Bhalla K, et al.Prevention of apoptosis by bcl-2: release of cytochrome C from motichondria,[J].Science, 1997; 275: 1129—1132.
4. Adams JM,Copy S.The Bcl-2 protein familyrarbiters of cell survival[J].Science,1998,281:1332—1336.
5. Boise LH.Garcia MG,Postems CE,et al.Bcl-x, a bcl-2-related gene that function as a dominant regulator of apoptosis cell death. Cell,1993,74:597-608.
6. Shimizn S, Eguchi Y,Kosada H,et al .Prevention of Hypoxia-induced cell death by Bcl-2 and bcl-XL. Nature,1995,374:811-813.
7. Cheng EH,Kirsch DC,Clem RJ,et al.Convension of Bcl-2 to Bax like death effector by Caspase[J]. Science,1997,278:1966-1968.
8. Wright ME, Han DK, Hockenbery DM. Caspase-3 and inhibitor of apoptosis protein interaction in saccharomyces cerevisiae and mammalian cells.FEBS Lett, 2000, 481:13—14.
9. Yigong Shi.Mechenisms of caspase activation and inhibition during apoptosis[J]. Mol Cell, 2002,9:459-470.
10. Raff M.Cell suicide for beginners[J].Nature,1998,396:119—122.
11. Fridlander RM, Yuan J .ICE,neuronal apoptosis and neurodegeneration[J].Cell Death Differ,1998,5(10):823—831.
12. Li P,Nijihawan D,Budihardjo L,et al.Cytochrome C and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptosis protease casade.[J].Cell, 1997,91:479—489.
13. Ahonad M,Srinivasula SM,Hegde R,et al. Identification and characterication of murine caspase-14,a new member of the caspase family.CancerRes,1998,58:5201-5205.
14. Li H,Zhu H,Xu C,et al. Cleavge of Bid by bcl-2 :release by caspase-8 mediates the mitochondrial damage in the Fas pathway of apoptosis.[J]. Cell, 1998,94:491—501.
15. Budihardjo I,Oliver H,Lutter M,et al.Biochemical of Bid by caspase-8 mediates the mitochondrial damage in the Fas pathway of apoptosis[J].Annu Rev Cell Del Bio,1999,15: 269—290.
16. Askenazi A, Dixit VM.Death receptors signaling and modulation[J].Science,1998,281:1305—1308.
17. Nakagawa T,Zhu H,Morishima N, et al.Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotixicity amyloid[J].Nature,2000,403:98-103.
18. Rammohan VR,Evan et al.Coupling endoplasmic-reticulum stress to cell death program[J]. J Biol Chem,2001,276:3869-3874.
19. Vilen A, Movsesyan G, et al.Ceramide induces neuronal apoptosis throughpathway. Biochemical Research Communications.Volume 299, Issue2, 29 November 2002, pages 201—207.
20. Kondo S,Bama Bp,Morimura T,et al.Interleukin-1 B coverting enzyme mediates cisplatin-induced apoptosis in malignant glioma cells[J].Cancer Res, 1995,55:6166—6171.
2 Reed JC. Dysregulation of apoptosis in cancer[J].J Chin Oncol, 1999, 17: 2941—2953。
3 Hengarter MO. The biochemistry of apoptosis[J]. Nature, 2000, 407: 770-776.
4 Yang J, Liu XS, Bhalla K, et al. Science, 1997; 275: 1129—1132.
5 Adams JM, Copy S. The Bcl-2 protein family: arbiters of cell survival[J]. Science, 1998, 281: 1332—1336.
6 Wright ME, Han DK, Hockenbery DM. Caspase-3 and inhibitor of apoptosis protein interaction in saccharomyces cerevisiae and mammalian cells. FEBS Lett, 2000, 481: 13—14.
7 Deveraux Q. L., Reed J. C. IAP family proteins-suppressors of apoptosis [J]. Gene Develp, 1999; 13 (3): 239—252.
8 Raff M. Cell suicide for beginners[J]. Nature, 1998, 396(6707): 119—122.
9 Fridlander RM, Yuan J.ICE, neuronal apoptosis and neurodegeneration[J]. Cell Death Differ, 1998, 5(10): 823—831.
10 Li P, Nijihawan D, Budihardjo L, et al. Cytochrome C and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptosis protease casade. [J]. Cell, 1997, 91: 479—489.
11 Budihardjo I, Oliver H, Lutter M, et al. Biochemical of Bid by caspase-8 mediates the mitochondrial damage in the Fas pathway of apoptosis[J]. Annu Rev Cell Del Bio, 1999, 15: 269—290.
12 Askenazi A, Dixit VM, Death receptors: signaling and modulation[J]. Science, 1998, 281: 1305—1308.
13 Hu S, Snipas SJ, Vincenz C, et al. Caspase-14 is a novel developmentally regulated protease[J]. J Biol Chem, 1998, 273: 29648—29653.
14 Hirata H, Takahashi A, Kobayashi S, et al. Caspases are activated in a branched protease cascade and control distinct downstream processes in Fas-induced apoptosis[J]. J Exp Med, 1998, 187(4): 587—600.
15 Stennicke HR, Salvesen GS. Properties of the caspase[J]. Bichim Biophys Acta, 1998, 1387: 17—31.
16 中华病理学杂志编辑委员会，《全国免疫组织化学技术与诊断标准化专题研讨会》会议记要[J]．中华病理学杂志，1996；25(6)：326．
17 徐建国．颅咽管瘤的动物模型的建立及肿瘤生物学特性的研究．四川大学临床医学博士专业学位论文．B202658．2005，4．
18 Schut, Louis MD, Discussant, et al. Management of a Pediatric Hypothalamic Mass. [Miscellaneous Article]. Neurosurgery. 1996, Apr, 38(4): 806-811.
19 Liu JM, Garonzik IM, Eberhart CG, et al. Ectopic recurrence of craniopharygioma after an interhemispheric transcallosal approacha: case report. [Revew]. Neurosurgery. 2002 Mar, 50(3): 639-644; discussion644-645.
20 Krupp W, Geiger K, Schober R, et al. Cytogenetic and molecular cytogenetic analyses in diffuse astrocytomas [J]. Cancer Genet Cytogenet, 2004Aug: 153 (1): 32——38.
21 Krajewaka M, Wang HG; Krajewaki S, et al. Immunohistochemistry of in vivo patterns of expression of CPP32(caspase3),a cell death protease. Cancer Res, 1997, 57: 1605—1613.
22 孙宝华，张俊，王保菊等．Caspase3基因表达与原发性肝细胞癌凋亡 无关．中华肿瘤杂志．2000，22(1)39—41．
23 文亚渊，刘宝华．基因Survivin、Caspase-3在胃癌发生中的作用．消化外科，2004，3(6)：415—418．
24 文亚渊，刘宝华，付涛．基因Survivin、Caspase-9在胃癌组织中的表达及与胃癌发生的关系．中国肿瘤临床，2005，32(17)：969—972．
25 高红，王常林，蔡炜嵩等．Survivin、Caspase-3mRNA在肾母细胞瘤中的表达及意义．中国当代儿科杂志，2004，6(5)：381—384．
26 周舟，王小华，朱光旭等．Caspase-3、9表达上调参与缺氧诱导心肌细胞凋亡．第三军医大学学报，2005，27(3)：185-188．
27 徐强，司良毅，赵小兰等．大鼠心肌缺血再灌注时心肌细胞凋亡caspase-3活性变化规律及药物对其影响．第三军医大学学报，2003，25(15)：1335-1357．
28 易成腊，陈安民，徐卫国等．大鼠脊髓损伤后Caspase-3表达和神经细胞凋亡相关性研究．创伤外科杂志．2004，6(3)：171—174．
29 Springer JE, Azbil RD, Knapp PE. Activation of the Caspase-3 apoptotic cascade in traumatic spinal cord injury[J]. Nat Med, 1999, 5(8): 943—946.
30 Cheng Y, Deshmukh M, Dcosta A, et al. Caspase inhibitor affords neuroprotection with delayed administration in a rat model of neonatal hypoxicischemic brain injury. J clin invest, 1998, 101(9): 1992.
31 郭燕舞，杨鹏飞，徐如祥．Caspase-3在脑损伤时的作用．广东医学．2004，25(9)：1026—1028．
32 James J, Velier A, Ellison K, et al. Caspase-8 and Caspase-3 Are Expressed by Different Populations of Cortical Neurons Undergoing Delayed Cell Death after Focal Stroke in the Rat. The Journal of Neuroscience, July 15, 1999, 19(14): 5932—5941.
33 郭云良，陈红兵，高英茂．大鼠脑缺血再灌注后神经细胞凋亡与Caspase-3和Caspase-9基因表达．解剖学杂志．2003，26(5)：430—434．
34 Gong C, Boulis N, Qian J, et al. Intracerebral hemorrhage-induced neuronal death. Neurosurgy. 2001, 48 (4): 875.
35 吴家幂，李珺．Caspase-3在脑出血后神经细胞凋亡中的作用．放射免疫学杂志．2005，18(2)：135—137．
36 Emery E, Aldana P, Bunge MB, et al. Apoptosis after traumatic human spinal cord injury[J]. J Neurosurg, 1998, 89(6): 911—920.
37 王文昭，戚基萍，王得升等．人脑出血后海马神经元损伤与Caspase-3的关系．中风与神经疾病杂志．05．22(2)：107—110．
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39 杨建青，黄柏英，吕新生．Caspase-9在氟尿嘧啶诱导的肝癌细胞凋亡中的作用．中国普通外科杂志．2003，12(3)：173—175．
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