Bcl-2和Fas/FasL在胆道肿瘤中的相关性研究
摘要
目的：研究Bcl-2和Fas／FasL在胆囊癌组织的表达及其相关性，并探讨其生物学意义。方法：应用免疫组化法研究33例胆囊癌，19例胆囊腺瘤，3例胆囊上皮不典型增生和20例慢性胆囊炎组织中Bcl-2蛋白和Fas／FasL蛋白的表达水平及特征。检测了胆管癌细胞株QBC939细胞和人肝门部胆管癌组织中FasL的表达；并用TUNEL技术检测了相关淋巴细胞的凋亡。结果：Bcl-2蛋白的阳性率在胆囊癌、胆囊腺瘤和胆囊上皮不典型增生分别为78.8％(26／33)、73.7％(14／19)和100％(3／3)，三者间无显著差异；而Bcl-2蛋白在胆囊癌和胆囊腺瘤的阳性率均显著高于慢性胆囊炎组织的阳性率40％(8／20)(P＜0.05)。FasL蛋白的阳性率在胆囊癌、胆囊腺瘤和胆囊上皮不典型增生分别为81.8％(27／33)、84.2％(16／19)和100％(3／3)，三者间无显著差异；而FasL蛋白在胆囊癌和胆囊腺瘤的阳性率均显著高于慢性胆囊炎组织的阳性率55％(11／20)(P＜0.05)。Bcl-2蛋白和FasL蛋白的阳性率均与其分化程度和Nevin分期无关。Fas蛋白的阳性率在慢性胆囊炎、胆囊腺瘤和胆囊上皮不典型增生分别为80％(16／20)、73.7％(14／19)和100％(3／3)，三者间无显著差异；而Fas蛋白在胆囊癌的阳性率为39.4％(13／33)，显著低于在慢性胆囊炎和胆囊腺瘤的阳性率(P＜0.05)；Fas蛋白在低分化胆囊癌的阳性率为25％(5／20)，显著低于在高-中分化胆囊癌的阳性率61.5％(8／13)(P＜0.05)，但与Nevin分期无关。Bcl-2蛋白和FasL蛋白在胆囊癌的表达呈显著正相关(r_s=0.478，P＜0.005)，Bcl-2蛋白和Fas蛋白在胆囊癌的表达呈显著负相关(r_s=-0.316，P＜0.05)，Fas蛋白和FasL蛋白在胆囊癌的表达无显著相关性。在所观察的手术切除的肝门部胆管癌组织中均发现FasL蛋白的表达，TUNEL染色发现瘤周肿瘤浸润淋巴细胞(TIL)有高水平的细胞死亡；在胆管癌细胞中检测到FasL蛋白的表达。结论：Bcl-2蛋白和Fas／FasL蛋白的表达与胆囊癌的生物学行为相关，且具有显著的相互协同作用。胆管癌可通过FasL诱导活化淋巴细胞的凋亡以逃避免疫监视。
Objective: To study the expression of Bcl-2 protein and Fas/FasL protein in gallbladder carcinoma, and their biological behaviors and correlation. And to assess its ability to induce apoptosis and immune escape. Methods: Avidin-biotin-peroxidase complex method was used to study the expression of Bcl-2 and Fas/FasL in 33 cases of gallbladder carcinoma, 19 cases of gallbladder adenoma, 3 cases of gallbladder epithelial dysplasia and 20 cases of chronic cholecystis. The expression of FasL in 48 specimens of human hilar cholangiocacinoma resected in operation and cultured QBC939 cholangiocarcinoma cell line was detected by immunohistochemistry. TUNEL was used to detect apoptotic cells. Results: The positive rates of Bcl-2 in carcinoma, adenoma and dysplasia were 78.8%(26/33), 73.7%(14/19) and 100%(3/3) respectively, there were not significant difference among them. The positive rates of Bcl-2 in cholecystis was 40%(8/20). The positive rates of Bcl-2 in carcinoma and adenoma were significantly higher than in cholecystis (p<0.05): The positive rates of FasL in carcinoma, adenoma and dysplasia were 81.8%(27/33), 84.2%(16/19) and 100%(3/3) respectively, there were not significant difference among them. The positive rates of FasL in cholecystis were 55%(11/20). The positive rates of FasL in carcinoma and adenoma were significantly higher than in cholecystis (p<0.05). The expression of Bcl-2 and FasL were not correlated with metastasis and differentiation. The positive rates of Fas in cholecystis, adenoma and dysplasia were 80%(16/20), 73.7%(14/19) and 100%(3/3) respectively, there were not significant difference among them. The positive rates of Fas in carcinoma were 39.4%(13/33), which was significant lower than in cholecytis and adenoma (p<0.05). And the positive rates of Fas in poorly-differentiated carcinoma were significantly lower than in well-moderated-differentiated carcinoma (p<0.05), but it was not correlated with metastasis. The expression of FasL and Bcl-2 in gallbladder carcinoma took on some extent positive correlation(r_s=0.478, P<0.005), the expression of Fas and Bcl-2 in gallbladder carcinoma took on some extent negative correlation(r_s=-0.316, P<0.05), the expression of Fas and FasL in gallbladder carcinoma were not significant correlation. Prevalent expression of FasL was detected in 48 resected hilarcholangiocarcinomas examined. TUNEL staining disclosed a high level of cell death among lymphocytes infiltrating FasL positive areas of tumor. FasL protein was expressed in cholangiocarcinoma cells. Conclusions: The expression of Bcl-2 and Fas/FasL might be correlated with the biological behaviors of gallbladder carcinoma. And there might be some extent coordinated regulation among them. The hilar cholangiocarcinoma eludes immunological surveillance by inducing, via the Fas/ FasL system , the apoptosis of activated lymphocytes.
Objective: To study the expression of Bcl-2 protein and Fas/FasL protein in gallbladder carcinoma, and their biological behaviors and correlation. And to assess its ability to induce apoptosis and immune escape. Methods: Avidin-biotin-peroxidase complex method was used to study the expression of Bcl-2 and Fas/FasL in 33 cases of gallbladder carcinoma, 19 cases of gallbladder adenoma, 3 cases of gallbladder epithelial dysplasia and 20 cases of chronic cholecystis. The expression of FasL in 48 specimens of human hilar cholangiocacinoma resected in operation and cultured QBC939 cholangiocarcinoma cell line was detected by immunohistochemistry. TUNEL was used to detect apoptotic cells. Results: The positive rates of Bcl-2 in carcinoma, adenoma and dysplasia were 78.8%(26/33), 73.7%(14/19) and 100%(3/3) respectively, there were not significant difference among them. The positive rates of Bcl-2 in cholecystis was 40%(8/20). The positive rates of Bcl-2 in carcinoma and adenoma were significantly higher than in cholecystis (p<0.05): The positive rates of FasL in carcinoma, adenoma and dysplasia were 81.8%(27/33), 84.2%(16/19) and 100%(3/3) respectively, there were not significant difference among them. The positive rates of FasL in cholecystis were 55%(11/20). The positive rates of FasL in carcinoma and adenoma were significantly higher than in cholecystis (p<0.05). The expression of Bcl-2 and FasL were not correlated with metastasis and differentiation. The positive rates of Fas in cholecystis, adenoma and dysplasia were 80%(16/20), 73.7%(14/19) and 100%(3/3) respectively, there were not significant difference among them. The positive rates of Fas in carcinoma were 39.4%(13/33), which was significant lower than in cholecytis and adenoma (p<0.05). And the positive rates of Fas in poorly-differentiated carcinoma were significantly lower than in well-moderated-differentiated carcinoma (p<0.05), but it was not correlated with metastasis. The expression of FasL and Bcl-2 in gallbladder carcinoma took on some extent positive correlation(r_s=0.478, P<0.005), the expression of Fas and Bcl-2 in gallbladder carcinoma took on some extent negative correlation(r_s=-0.316, P<0.05), the expression of Fas and FasL in gallbladder carcinoma were not significant correlation. Prevalent expression of FasL was detected in 48 resected hilarcholangiocarcinomas examined. TUNEL staining disclosed a high level of cell death among lymphocytes infiltrating FasL positive areas of tumor. FasL protein was expressed in cholangiocarcinoma cells. Conclusions: The expression of Bcl-2 and Fas/FasL might be correlated with the biological behaviors of gallbladder carcinoma. And there might be some extent coordinated regulation among them. The hilar cholangiocarcinoma eludes immunological surveillance by inducing, via the Fas/ FasL system , the apoptosis of activated lymphocytes.
引文
[1] Carson DA, Ribeiro JM. Apoptosis and disease. Lancet, 1993, 341(8855): 1251-4.
[2] 王少峰，洪道显等．大肠腺瘤—癌序列中p53和bcl—2蛋白的表达及其与细胞凋亡的关系．中华消化杂志，2002(9)：564—565．
[3] Weller M, Malipiero U, Aguzzi A, et al. Protooncogene bcl-2 gene transfer abrogates Fas/APO-1 antibody-mediated apoptosis of human malignant glioma cells and confers resistance to chemotherapeutic drugs and therapeutic irradiation. J Clin Invest, 1995,95(6):2633-43.
[4] 龚非力．医学免疫学．第一版版．北京：科学出版社，2000．261．
[5] 范毓东．Bcl-2与胆管细胞癌．国外医学外科学分册，1999，26(3)：159-161．
[6] Tsujimoto Y, Finger LR, Yunis J, et al. Cloning of the chromosome breakpoint of neoplastic B cells with the t(14;18) chromosome translocation. Science, 1984,226(4678): 1097-9.
[7] 谷化平，尚培中，周翠玲．胆囊癌组织CD15和Bcl-2的表达及相关性研究．临床肝胆病杂志，2001，17(1)：57-58．
[8] Frei H. Cancer Medicine. 5th edition ed. Singapore:Harcourt Asia Pte Ltd, 2001.62.
[9] Tsujimoto Y, Croce CM. Analysis of the structure, transcripts, and protein products of bcl-2, the gene involved in human follicular lymphoma. Proc Natl Acad Sci U S A, 1986,83(14):5214-8.
[10] Reed JC. Double identity for proteins of the Bcl-2 family. Nature, 1997, 387 (6635):773-6.
[11] Green DR. Apoptotic pathways: the roads to ruin. Cell, 1998,94(6):695-8.
[12] Mignotte B, Vayssiere JL. Mitochondria and apoptosis. Eur J Biochem, 1998, 252(1):1-15.
[13] Green DR, Reed JC. Mitochondria and apoptosis. Science, 1998,281(5381): 1309-12.
[14] Frei H. Cancer Medicine. 5th edition ed. Singapore:Harcourt Asia Pte Ltd, 2001.1361.
[15] McDonnell TJ, Troncoso P, Brisbay SM, et al. Expression of the protooncogene bcl-2 in the prostate and its association with emergence of androgen-independent prostate cancer. Cancer Res, 1992,52(24):6940-4.
[16] 姜海行，聂海明，邓德海等．幽门螺杆菌体外诱导大鼠胃黏膜上皮细胞凋亡．世界华人消化杂志，2005，13(24)：2838-2841．
[17] 董来华，马君红，茅育蕾等．胃黏膜异型增生c-myc、bcl-2、P53的表达及其意义．中华消化内镜杂志，2003，(20)5：325-327．
[18] 叶定伟，李慧等．雄激素对凋亡抑制蛋白Bcl—XL表达的影响．中华泌尿外科杂志．2001，22(4)：248-248．
[19] Harnois DM, Que FG, Celli A, et al. Bcl-2 is overexpressed and alters the threshold for apoptosis in a cholangiocarcinoma cell line. Hepatology, 1997, 26(4):884-90.
[20] Fiorentino M, D'Errico A, Altimari A, et al. High levels of 13CL-2 messenger RNA detected by in situ hybridization in human hepatocellular and cholangiocellular carcinomas. Diagn Mol Pathol, 1999,8(4): 189-94.
[21] Mikami T, Yanagisawa N, Baba H, et al. Association of Bcl-2 protein expression with gallbladder carcinoma differentiation and progression and its relation to apoptosis. Cancer, 1999,85(2):318-25.
[22] 夏金荣，赵东强．细胞凋亡及其信号转导、凋亡抵抗与恶性肿瘤．临床荟萃．2005，20(24)：1421-1424．
[23] 程磊 王跃．细胞凋亡与胃癌．国外医学：临床生物化学与检验学分册．2004年25卷3期：265-268．
[24] Schulze-Osthoff K, Ferrari D, Los M, et al. Apoptosis signaling by death receptors. Eur J Biochem, 1998,254(3):439-59.
[25] Shimonishi T, Isse K, Shibata F, et al. Up-regulation of fas ligand at early stages and down-regulation of Fas at progressed stages of intrahepatic Cholangiocarcinoma reflect evasion from immune surveillance. Hepatology, 2000,32(4 Pt 1):761-9.
[26] Griffith TS, Ferguson TA. The role of FasL-induced apoptosis in immune privilege. Immunol Today, 1997,18(5):240-4.
[27] 李佳莉，赵丽纯．Fas／FasL系统与肿瘤的免疫逃逸．国外医学免疫学分册，2001，24(2)：65-67．
[28] O'Connell J, O'Sullivan GC, Collins JK, et al. The Fas counterattack: Fas-mediated T cell killing by colon cancer cells expressing Fas ligand. J Exp Med, 1996,184(3):1075-82.
[29] 马永会，刘云会，王文军等．Fas／FasL与肿瘤免疫逃逸．沈阳医学院学报．2003，5(3)：199-201．
[30] 张煊．肿瘤免疫逃逸机制研究新进展．中国肿瘤生物治疗杂志．2001，8(1)：72-74．
[31] Nagata S, Golstein P. The Fas death factor. Science, 1995,267(5203): 1449-56.
[32] Griffith TS, Brunner T, Fletcher SM, et al. Fas ligand-induced apoptosis as a mechanism of immune privilege. Science, 1995,270(5239):1189-92.
[33] Bellgrau D, Gold D, Selawry H, et al. A role for CD95 ligand in preventing graft rejection. Nature, 1995,377(6550):630-2.
[34] Que FG, Phan VA, Phan VH, et al. Cholangiocarcinomas express Fas ligand and disable the Fas receptor. Hepatology, 1999,30(6):1398-404.
[35] Wang SG, Han BL, Duan HC, et al. Establishment of a cell line ofextrahepatic bile duct carcinoma. Chin J Exp Surg, 1997,14:67-68.
[36] Stuart PM, Griffith TS, Usui N, et al. CD95 ligand (FasL)-induced apoptosis is necessary for corneal allograft survival. J Clin Invest, 1997,99(3):396-402..
[37] Zhang H, Yang Y, Horton JL, et al. Amelioration of collagen-induced arthritis by CD95 (Apo-1/Fas)-ligand gene transfer. J Clin Invest, 1997,100(8): 1951-7.
[1] Weller M, Maliplero U, Aguzzi A, et al. Protooncogene bcl-2 gene transfer abrogates fas/apo-1 antib0dy-mediated apoptosis of human malignant glioma cells and confers resistance to chemotherapeutic drugs and therapeutic irradiation. J Clin Invest, 1995, 95:2633-2643
[2] Griffith TS, Fergnson TA. The role of fasL-induced apoptosis in immune privilege. Immunol Today, 1997, 18(5):240
[3] 龚非力．医学免疫学，第一版．北京：科学版社，2000：261
[4] Schulze-Osthoff K, Ferrari D, Los M, et al. Apoptosis signaling by death receptors. Eur J Biochem, 1998, 254:439-459
[5] 李佳莉，赵丽纯．Fas／FasL系统与肿瘤的免疫逃逸．国外医学免疫学分册，2001，24(2)：65-67
[6] Gdffith TS, Brenner T, Fletcher SM, et al. Fas ligand-induced apoptosis as a mechanism of immune privilege. Science, 1995, 270(17): 1189-1192
[7] Bellgrau D, Gold D, Selawry H, et al. A role for CD95 ligand in preventing graft rejection.Nature, 1995,377(19):630-632
[8] Suda T, Takahashi T, Golstein P, et al. Molecular cloning and expressing of the fas ligand, a novel member of the tumor necrosis factor family.Cell, 1993, 75(17):1169-1178
[9] French LE, Hahne M, Viard I, et al. Fas and fas ligand in embryos and adult mice:ligand expression in several immune-privileged tissues and coexpression in adult tissues characterized by apoptotic cell turnover. J Cell Biol, 1996, 133(2):335-343
[10] Frei H. Cancer Medicine, 5th edition. Singapore: Harcourt Asia Pte Ltd, 2001: 24-28
[11] Shimonishi T, Isse T, Shibata F, et al. Up-regulation of fas ligand at early stages and down-regulation of fas at progressed stages of intrahepatic cholangiocarcinoma reflect evasion from immune surveillance. Hepatology, 2000, 32:761-769
[12] Cohen JJ. Apoptosis: Mechanisms of life and death in the immune system. J Allergy Clin Immunol, 1999, 4:548-554
[13] Lowin B, Hahne M, Mattmann C, et al. Cytolytic T-cell cytotoxicity is mediated through perfodn and fas lytic pathways. Nature, 1994, 370:650-652
[14] Ashkenazi A, Dixit VM. Death receptors: signaling and modulation. Science, 1998, 281:1305-1308
[15] Walczak E, Miller RE, Ariail K, et al. Tumoricidal activity oftumor necrosis factor-related apoptosis-inducing ligand in vivo. Nature Medicine, 1999, 5:157-163
[16] O'Connell J, O'Sullivan GC, Collins JK, et al. The fas counterattack: fasmediated T cell killing by colon cancer cells expressing fas ligand. J Exp Med, 1996, 184:1075-1082
[17] Hahne M, Rimoldi D, Schroter M, et al. Melanoma cell expression of fas(apo-1/CD95) ligand:implications for tumor immune escape. Science, 1996, 274(22): 1363-1366
[18] Niehans GA, Rrnnner T, Frizelle SP, et al. Human lung carcinomas express fas ligand. Cancer Res, 1997, 57:1007-1012
[19] Saas P, Walker PR, Hahne M, et al. Fas ligand expression by astrocytoma in vivo:maintaining immune privilege in the brain?. J Clin Invest, 1997, 99(6):1173-1178
[20] Bennett MW, O'Connell J, O'Sullivan GC, et al. Expression of fas ligand by human gastric adenocarcinomas: a potential mechanism of immune escape in stomach cancer. Gut, 1999, 44:156-162
[21] Keane MM, Ettenberg SA, Lowrey GA, et al. Fas expression and function in normal and malignant breast cell lines. Cancer Res, 1996, 56:4.791-4798
[22] Walker PR, Saas P, Dietrich PY, et al. Role of fas ligand(CD95L) in immune escape. J Immunol, 1997, 158:4521-4524
[23] 范毓东．Bcl-2与胆管细胞癌．国外医学外科学分册，1999，26(3)：159-161
[24] Tsujimoto Y, Finger LR, Yunis J, et al. Cloning of the chromosome breakpoint neoplastic B cells with the t(14; 18) chromosome translocation. Science, 1984, 226(30):1097-1099
[25] 谷化平，尚培中，周翠玲．胆囊癌组织CD15和Bcl-2的表达及相关性研究．临床肝胆病杂志，2001，17(1)：57-58
[26] Frei H. Cancer Medicine, 5th edition. Singapore: Harcourt Asia Pte Ltd, 2001: 62
[27] Tsujimoto Y, Croce CM. Analysis of the structure, transcripts, and protein products of bcl-2, the gene involved in human follicular lymphoma. Proc Natl Acad Sci USA, 1986, 83:5214-5218
[28] Carson DA, Ribeiro JM. Apoptosis and disease. The lancet, 1993, 341:1251-1254
[29] Reed JC. Double identity for proteins of the bcl-2 family. Nature, 1997, 387:773-776
[30] Green DR. Apoptotic pathways: The roads to ruin. Cell, 1998, 94:695-698
[31] Mignotte B, Vayssiere JL. Mitochondria and apoptosis. Eur. J. Biochem, 1998, 252:1-15
[32] Green DR, Reed JC. Mitochondria and apoptosis. Science, 1998, 281:1309-1312
[33] Frei H. Cancer Medicine, 5th edition. Singapore: Harcourt Asia Pte Ltd, 2001: 1361
[34] McDonnell TJ, Troncoso P, Brisbay SM, et al. Expression of the protooncogene bcl-2 in the prostate and its association with emergence of androgen-independent prostate cancer. Cancer Res, 1992,52:6940-6944
[35] Hamois DM, Que FG, Celli A, et al. Bcl-2 is overexpressed and alters the threshold for apoptosis in a cholangiocarcinoma cell line. Hepatology, 1997, 26:884-890
[36] Fiorentino M, D'Erdco AD, Altimari A, et al. High levels of bcl-2 messenger RNA detected by in situ hybridization in human hepatocellular and cholangiocellular carcinomas. Diagn Mol Pathol, 1999, 8(4): 189-194
[37] Mikami T, Yanagisawa N, Baba H, et al. Association of bcl-2 protein expression with gallbladder carcinoma differentiation and progression and its relation to apoptosis. Cancer, 1999, 85(2):318-325
[38] Itoh N, Tsujimoto Y, Nagata S. Effect of bcl-2 on fas antigen-mediated cell death. J Immunol, 1993, 151(2):621-627
[39] Yoshino T, Kondo E, Cao L, et al. Inverse expression ofbcl-2 protein and fas antigen in lymphoblasts in peripheral lymph nodes and activated peripheral blood T and B lymphocytes. Blood, 1994, 83(7):1856-1861
[40] Mandal M, Maggirwar SB, Sharma N, et al. Bcl-2 prevents CD95 (fas/apo-1)-induced degradation of lamin B and poly(ADP-ribose) polymerase and restores the NF-κ B signaling pathway. J Biol Chem, 1996, 271(48):30354-30359
[41] Memon SA, Moreno MB, Petrak D, et al. Bcl-2 blocks glucocorticoid-but not fas-or activation-induced apoptosis in a T cell hybddoma. J Immunol, 1995, 155:4644-4652
[42] 杨光华．病理学，第五版．北京：人民卫生出版社，2001：113
[43] 吴在德．外科学，第五版．北京：人民卫生出版社，2000：248
[2] 王少峰，洪道显等．大肠腺瘤—癌序列中p53和bcl—2蛋白的表达及其与细胞凋亡的关系．中华消化杂志，2002(9)：564—565．
[3] Weller M, Malipiero U, Aguzzi A, et al. Protooncogene bcl-2 gene transfer abrogates Fas/APO-1 antibody-mediated apoptosis of human malignant glioma cells and confers resistance to chemotherapeutic drugs and therapeutic irradiation. J Clin Invest, 1995,95(6):2633-43.
[4] 龚非力．医学免疫学．第一版版．北京：科学出版社，2000．261．
[5] 范毓东．Bcl-2与胆管细胞癌．国外医学外科学分册，1999，26(3)：159-161．
[6] Tsujimoto Y, Finger LR, Yunis J, et al. Cloning of the chromosome breakpoint of neoplastic B cells with the t(14;18) chromosome translocation. Science, 1984,226(4678): 1097-9.
[7] 谷化平，尚培中，周翠玲．胆囊癌组织CD15和Bcl-2的表达及相关性研究．临床肝胆病杂志，2001，17(1)：57-58．
[8] Frei H. Cancer Medicine. 5th edition ed. Singapore:Harcourt Asia Pte Ltd, 2001.62.
[9] Tsujimoto Y, Croce CM. Analysis of the structure, transcripts, and protein products of bcl-2, the gene involved in human follicular lymphoma. Proc Natl Acad Sci U S A, 1986,83(14):5214-8.
[10] Reed JC. Double identity for proteins of the Bcl-2 family. Nature, 1997, 387 (6635):773-6.
[11] Green DR. Apoptotic pathways: the roads to ruin. Cell, 1998,94(6):695-8.
[12] Mignotte B, Vayssiere JL. Mitochondria and apoptosis. Eur J Biochem, 1998, 252(1):1-15.
[13] Green DR, Reed JC. Mitochondria and apoptosis. Science, 1998,281(5381): 1309-12.
[14] Frei H. Cancer Medicine. 5th edition ed. Singapore:Harcourt Asia Pte Ltd, 2001.1361.
[15] McDonnell TJ, Troncoso P, Brisbay SM, et al. Expression of the protooncogene bcl-2 in the prostate and its association with emergence of androgen-independent prostate cancer. Cancer Res, 1992,52(24):6940-4.
[16] 姜海行，聂海明，邓德海等．幽门螺杆菌体外诱导大鼠胃黏膜上皮细胞凋亡．世界华人消化杂志，2005，13(24)：2838-2841．
[17] 董来华，马君红，茅育蕾等．胃黏膜异型增生c-myc、bcl-2、P53的表达及其意义．中华消化内镜杂志，2003，(20)5：325-327．
[18] 叶定伟，李慧等．雄激素对凋亡抑制蛋白Bcl—XL表达的影响．中华泌尿外科杂志．2001，22(4)：248-248．
[19] Harnois DM, Que FG, Celli A, et al. Bcl-2 is overexpressed and alters the threshold for apoptosis in a cholangiocarcinoma cell line. Hepatology, 1997, 26(4):884-90.
[20] Fiorentino M, D'Errico A, Altimari A, et al. High levels of 13CL-2 messenger RNA detected by in situ hybridization in human hepatocellular and cholangiocellular carcinomas. Diagn Mol Pathol, 1999,8(4): 189-94.
[21] Mikami T, Yanagisawa N, Baba H, et al. Association of Bcl-2 protein expression with gallbladder carcinoma differentiation and progression and its relation to apoptosis. Cancer, 1999,85(2):318-25.
[22] 夏金荣，赵东强．细胞凋亡及其信号转导、凋亡抵抗与恶性肿瘤．临床荟萃．2005，20(24)：1421-1424．
[23] 程磊 王跃．细胞凋亡与胃癌．国外医学：临床生物化学与检验学分册．2004年25卷3期：265-268．
[24] Schulze-Osthoff K, Ferrari D, Los M, et al. Apoptosis signaling by death receptors. Eur J Biochem, 1998,254(3):439-59.
[25] Shimonishi T, Isse K, Shibata F, et al. Up-regulation of fas ligand at early stages and down-regulation of Fas at progressed stages of intrahepatic Cholangiocarcinoma reflect evasion from immune surveillance. Hepatology, 2000,32(4 Pt 1):761-9.
[26] Griffith TS, Ferguson TA. The role of FasL-induced apoptosis in immune privilege. Immunol Today, 1997,18(5):240-4.
[27] 李佳莉，赵丽纯．Fas／FasL系统与肿瘤的免疫逃逸．国外医学免疫学分册，2001，24(2)：65-67．
[28] O'Connell J, O'Sullivan GC, Collins JK, et al. The Fas counterattack: Fas-mediated T cell killing by colon cancer cells expressing Fas ligand. J Exp Med, 1996,184(3):1075-82.
[29] 马永会，刘云会，王文军等．Fas／FasL与肿瘤免疫逃逸．沈阳医学院学报．2003，5(3)：199-201．
[30] 张煊．肿瘤免疫逃逸机制研究新进展．中国肿瘤生物治疗杂志．2001，8(1)：72-74．
[31] Nagata S, Golstein P. The Fas death factor. Science, 1995,267(5203): 1449-56.
[32] Griffith TS, Brunner T, Fletcher SM, et al. Fas ligand-induced apoptosis as a mechanism of immune privilege. Science, 1995,270(5239):1189-92.
[33] Bellgrau D, Gold D, Selawry H, et al. A role for CD95 ligand in preventing graft rejection. Nature, 1995,377(6550):630-2.
[34] Que FG, Phan VA, Phan VH, et al. Cholangiocarcinomas express Fas ligand and disable the Fas receptor. Hepatology, 1999,30(6):1398-404.
[35] Wang SG, Han BL, Duan HC, et al. Establishment of a cell line ofextrahepatic bile duct carcinoma. Chin J Exp Surg, 1997,14:67-68.
[36] Stuart PM, Griffith TS, Usui N, et al. CD95 ligand (FasL)-induced apoptosis is necessary for corneal allograft survival. J Clin Invest, 1997,99(3):396-402..
[37] Zhang H, Yang Y, Horton JL, et al. Amelioration of collagen-induced arthritis by CD95 (Apo-1/Fas)-ligand gene transfer. J Clin Invest, 1997,100(8): 1951-7.
[1] Weller M, Maliplero U, Aguzzi A, et al. Protooncogene bcl-2 gene transfer abrogates fas/apo-1 antib0dy-mediated apoptosis of human malignant glioma cells and confers resistance to chemotherapeutic drugs and therapeutic irradiation. J Clin Invest, 1995, 95:2633-2643
[2] Griffith TS, Fergnson TA. The role of fasL-induced apoptosis in immune privilege. Immunol Today, 1997, 18(5):240
[3] 龚非力．医学免疫学，第一版．北京：科学版社，2000：261
[4] Schulze-Osthoff K, Ferrari D, Los M, et al. Apoptosis signaling by death receptors. Eur J Biochem, 1998, 254:439-459
[5] 李佳莉，赵丽纯．Fas／FasL系统与肿瘤的免疫逃逸．国外医学免疫学分册，2001，24(2)：65-67
[6] Gdffith TS, Brenner T, Fletcher SM, et al. Fas ligand-induced apoptosis as a mechanism of immune privilege. Science, 1995, 270(17): 1189-1192
[7] Bellgrau D, Gold D, Selawry H, et al. A role for CD95 ligand in preventing graft rejection.Nature, 1995,377(19):630-632
[8] Suda T, Takahashi T, Golstein P, et al. Molecular cloning and expressing of the fas ligand, a novel member of the tumor necrosis factor family.Cell, 1993, 75(17):1169-1178
[9] French LE, Hahne M, Viard I, et al. Fas and fas ligand in embryos and adult mice:ligand expression in several immune-privileged tissues and coexpression in adult tissues characterized by apoptotic cell turnover. J Cell Biol, 1996, 133(2):335-343
[10] Frei H. Cancer Medicine, 5th edition. Singapore: Harcourt Asia Pte Ltd, 2001: 24-28
[11] Shimonishi T, Isse T, Shibata F, et al. Up-regulation of fas ligand at early stages and down-regulation of fas at progressed stages of intrahepatic cholangiocarcinoma reflect evasion from immune surveillance. Hepatology, 2000, 32:761-769
[12] Cohen JJ. Apoptosis: Mechanisms of life and death in the immune system. J Allergy Clin Immunol, 1999, 4:548-554
[13] Lowin B, Hahne M, Mattmann C, et al. Cytolytic T-cell cytotoxicity is mediated through perfodn and fas lytic pathways. Nature, 1994, 370:650-652
[14] Ashkenazi A, Dixit VM. Death receptors: signaling and modulation. Science, 1998, 281:1305-1308
[15] Walczak E, Miller RE, Ariail K, et al. Tumoricidal activity oftumor necrosis factor-related apoptosis-inducing ligand in vivo. Nature Medicine, 1999, 5:157-163
[16] O'Connell J, O'Sullivan GC, Collins JK, et al. The fas counterattack: fasmediated T cell killing by colon cancer cells expressing fas ligand. J Exp Med, 1996, 184:1075-1082
[17] Hahne M, Rimoldi D, Schroter M, et al. Melanoma cell expression of fas(apo-1/CD95) ligand:implications for tumor immune escape. Science, 1996, 274(22): 1363-1366
[18] Niehans GA, Rrnnner T, Frizelle SP, et al. Human lung carcinomas express fas ligand. Cancer Res, 1997, 57:1007-1012
[19] Saas P, Walker PR, Hahne M, et al. Fas ligand expression by astrocytoma in vivo:maintaining immune privilege in the brain?. J Clin Invest, 1997, 99(6):1173-1178
[20] Bennett MW, O'Connell J, O'Sullivan GC, et al. Expression of fas ligand by human gastric adenocarcinomas: a potential mechanism of immune escape in stomach cancer. Gut, 1999, 44:156-162
[21] Keane MM, Ettenberg SA, Lowrey GA, et al. Fas expression and function in normal and malignant breast cell lines. Cancer Res, 1996, 56:4.791-4798
[22] Walker PR, Saas P, Dietrich PY, et al. Role of fas ligand(CD95L) in immune escape. J Immunol, 1997, 158:4521-4524
[23] 范毓东．Bcl-2与胆管细胞癌．国外医学外科学分册，1999，26(3)：159-161
[24] Tsujimoto Y, Finger LR, Yunis J, et al. Cloning of the chromosome breakpoint neoplastic B cells with the t(14; 18) chromosome translocation. Science, 1984, 226(30):1097-1099
[25] 谷化平，尚培中，周翠玲．胆囊癌组织CD15和Bcl-2的表达及相关性研究．临床肝胆病杂志，2001，17(1)：57-58
[26] Frei H. Cancer Medicine, 5th edition. Singapore: Harcourt Asia Pte Ltd, 2001: 62
[27] Tsujimoto Y, Croce CM. Analysis of the structure, transcripts, and protein products of bcl-2, the gene involved in human follicular lymphoma. Proc Natl Acad Sci USA, 1986, 83:5214-5218
[28] Carson DA, Ribeiro JM. Apoptosis and disease. The lancet, 1993, 341:1251-1254
[29] Reed JC. Double identity for proteins of the bcl-2 family. Nature, 1997, 387:773-776
[30] Green DR. Apoptotic pathways: The roads to ruin. Cell, 1998, 94:695-698
[31] Mignotte B, Vayssiere JL. Mitochondria and apoptosis. Eur. J. Biochem, 1998, 252:1-15
[32] Green DR, Reed JC. Mitochondria and apoptosis. Science, 1998, 281:1309-1312
[33] Frei H. Cancer Medicine, 5th edition. Singapore: Harcourt Asia Pte Ltd, 2001: 1361
[34] McDonnell TJ, Troncoso P, Brisbay SM, et al. Expression of the protooncogene bcl-2 in the prostate and its association with emergence of androgen-independent prostate cancer. Cancer Res, 1992,52:6940-6944
[35] Hamois DM, Que FG, Celli A, et al. Bcl-2 is overexpressed and alters the threshold for apoptosis in a cholangiocarcinoma cell line. Hepatology, 1997, 26:884-890
[36] Fiorentino M, D'Erdco AD, Altimari A, et al. High levels of bcl-2 messenger RNA detected by in situ hybridization in human hepatocellular and cholangiocellular carcinomas. Diagn Mol Pathol, 1999, 8(4): 189-194
[37] Mikami T, Yanagisawa N, Baba H, et al. Association of bcl-2 protein expression with gallbladder carcinoma differentiation and progression and its relation to apoptosis. Cancer, 1999, 85(2):318-325
[38] Itoh N, Tsujimoto Y, Nagata S. Effect of bcl-2 on fas antigen-mediated cell death. J Immunol, 1993, 151(2):621-627
[39] Yoshino T, Kondo E, Cao L, et al. Inverse expression ofbcl-2 protein and fas antigen in lymphoblasts in peripheral lymph nodes and activated peripheral blood T and B lymphocytes. Blood, 1994, 83(7):1856-1861
[40] Mandal M, Maggirwar SB, Sharma N, et al. Bcl-2 prevents CD95 (fas/apo-1)-induced degradation of lamin B and poly(ADP-ribose) polymerase and restores the NF-κ B signaling pathway. J Biol Chem, 1996, 271(48):30354-30359
[41] Memon SA, Moreno MB, Petrak D, et al. Bcl-2 blocks glucocorticoid-but not fas-or activation-induced apoptosis in a T cell hybddoma. J Immunol, 1995, 155:4644-4652
[42] 杨光华．病理学，第五版．北京：人民卫生出版社，2001：113
[43] 吴在德．外科学，第五版．北京：人民卫生出版社，2000：248