骨肉瘤组织中环氧合酶-2和P-耐药糖蛋白的表达及其相关性研究
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摘要
目的
骨肿瘤是威胁人类生命健康的疾病，尤以恶性骨肿瘤为甚。在我国良性骨肿瘤占骨肿瘤及瘤样病变发病的48.45％，发病年龄多发生在11-30岁，其中，男女发病率之比约为1.56:1.00，以骨软骨瘤为最多(31.60％)，其次为骨巨细胞瘤(23.96％)。恶性肿瘤占49.30％，发病年龄也多在11-30岁，骨肉瘤的发病率占恶性骨肿瘤的76.21％，居首位。当前，对骨肿瘤的治疗方法有很多种，其中包括外科手术，放疗，化疗，免疫疗法，中草药疗法等。其中化疗在骨肿瘤的治疗中的作用越来越重要，术前化疗，手术切除，术后化疗，这一治疗模式比单纯的手术切除要有效得多。而当今造成化疗失败的主要原因是产生化疗药物的耐药，肿瘤产生耐药的机制较为复杂，目前认为，肿瘤产生多药耐药最重要的分子机制是跨膜药泵基因的扩增或过表达，这类基因以MDR1(Mutidrug resistance-1)最为重要，其表达产物为一种膜相关糖蛋白-P-糖蛋白(P-glycoprotein，P-gp)。近年来研究发现，前列腺素(PGs)和一氧化氮(NO)在肿瘤的发展和血管生成过程中起着重要的作用。环氧合酶(COX)是前列腺素合成过程中的重要限速酶，环氧合酶-2(COX-2)的过度表达通过抑制凋亡、增加细胞外基质蛋白粘附、促进肿瘤血管生成等机制参与肿瘤的发生发展。本实验的研究目的是探讨环氧合酶-2(COX-2)和P-耐药糖蛋白(P-gp)在骨肉瘤组织标本中表达的临床意义及其相关性。
材料和方法
采用免疫组织化学方法(S-P法)检测31例骨肉瘤组织蜡块标本中COX-2和P-gp蛋白的表达情况，同时另取30例骨软骨瘤组织蜡块标本作为对照组，来对比它们之间的表达是否存在差异，并探讨其意义。
结果
骨肉瘤组织中COX一和P一gP蛋白的阳性表达率分别为61 .30%和58.
10%，显著高于对照组骨软骨瘤组织6.67%和10.00%(P<0.05)。COX-2
的表达与骨肉瘤组织学分级、肺转移有关(P=0.033，0.008)。P一gP的表达
与肺转移转移有关(P二0.022)，而与组织学分级关系不大(P二0.845)。
COX一的表达与P一gP的表达密切相关(r=0.398，P二0.027)。二者的表
达与患者的性别、年龄无关。
讨论
近年莱，根据流行病学的调查统计，非街体类抗炎药(NsAIDS)确实有
降低大肠癌发病率和死亡率的作用，而NSAIDs这一作用的发挥是通过对
环氧合酶一(cox一)的抑制效应而产生的。环氧合酶是前列腺素合成过程
中的重要的限速酶，目前发现，该酶有两种亚型，即COX一l和COX一。两者
虽然结构相似，但因其生物学特性上的差异，在正常组织和病理组织中发挥
着不同的作用。尤其在正常组织中不表达或是低表达的COX一基因，一旦
受到外界因素的刺激，就会被迅速而大量的诱导表达。本组研究资料显示，
31例骨肉瘤组织中，COX一阳性表达率为61 .30%，显著高于对照组骨软
骨瘤组织6.67%(P<0.05)。COX一的表达与骨肉瘤病变组织的组织学分
级呈正相关(P<0.05，P二0.033)。随着组织学分级的增高，COXZ的阳性
表达率增加。此外，COX一的表达与肺转移有关(P<0.05，P二0.008)。P-
gP通过其特有的”药泵”作用将化疗药物泵出肿瘤细胞，从而保护肿瘤细胞
免受化疗药物的细胞毒性作用。近年来的研究表明，P一gP的表达与肿瘤的
化疗敏感性和病人的生存时间呈负相关。本组P一gP的阳性表达率是58.
10%，明显高于骨软骨瘤组织10.00%(P<0.05)，P一gP的表达与肺转移有
关(P<0.05)。cox一和P一gP参与骨肉瘤的发生和发展过程，并且它们之
间具有协同作用。选择性的应用COX一抑制剂可能会预防肿瘤耐药的发
生，并且提高肿瘤化疗的疗效。
结论
在骨肉瘤中，COX一2、P一gP参与肿瘤的发生和发展过程，并且它们之间
具有协同作用。
Objective
The tumor of bone is a dangerous diseases for human, especially maliganant bone tumors, in our country ,it is 48.45% that the benignbone tumors account for the bone tumors and tumorous pathological changes,the age ,which the disease occurs, is from 11 to 30, man/woman is about 1. 56:1. 00, the osteochon-droma is the maximum ( 31. 60% ) , the second is the giant cell tumor of bone (23.96% ). it is 49. 30% that maliganant tumors account for the bone tumors and tumorous pathological changes,the age, which the disease occurs, is from 11 to 30, the osteosarcoma is the maximum(76. 21% ). nowadays,the methods of treating the tumor of bone are very many, including operation, chemic therapeutics , radiative therapeutics, immune therapeutics, therapeutics of Chinese traditional medicine, and so on . The chemic therapeutics is more and more important for the treatment of bone tumor . the mode, ( chemic therapeutics, operation , chemic therapeutics) is much effective than operation . but the reason that caused the chemi
c therapies failure is (drug resistance . the mechanism of the drug resistance is very complicated, now most think that the important mechanism of the drug resistance is that the gene of MDR1 overexpressed,itS outcome is a glycoprotein( P-gp). recent years,the reseaches have found that PCs and NO is very important for the development and the angiogenesis of tumor. COX is a pivotal limiting enzyme during the synthetical process of PCs . COX-2 takes part in the carcinogenesis and development of osteosarcoma by inhibiting apoptosis, enhancing the adhere of extracellular matrix and accelerating the angiogenesis of tumor, the aim of the experence is to investigate the expression of P-gp and COX-2 in osteosarcoma , as well to explore their relationship and clinical significance.
Materials and Methods
COX-2 and P-gp were detected by S-P immunohistochemical method in 31 cases of paraffin-embedded sections of osteosarcoma. at the same time, I took 30 cases of paraffin-embedded sections of osteochondroma as control groups, and conformed if there was difference between the two groups and discussed the significance.
Results
The positive rate of COX-2 and P-gp in human osteosarcoma was 61. 30% and 58. 10% , respectively. Significantly higher than osteochondroma (6. 67% , 10. 00% ). The expression of COX-2 was significantly associated with the histo-logical grades , lung metastasis ( P = 0. 033, 0. 008,). The expression of P-gp was significantly correlated with lung metastasis (P =0.022) , but not with his-tological grades ( P = 0. 845 ). There was a significant correlation between the expression of COX-2 and P-gp (r =0. 398,P =0.027). Conditions such as age,sex had no relation with the expression of COX-2 or P-gp.
Discussion
recent years,bases the epidemical survey, NSAIDS can strictly reduce the incidence and mortality of colon cancer, but NSAIDS operate by inhibiting COX-2. COX is a pivotal limiting enzyme during the synthetical process of PCs. by now, we have found that it has 2 subtypes,namely COX-1 and COX-2. they have similar structure but they have different biological characteristics, they play different functions in normal tissues and pathological tissues, in normal tissues, COX-2 gene does not express or express very lower. It will largely express as soon as it is activated by somethings. In my experience, The positive rate of COX-2 was 61. 30%. Significantly higher than osteochondroma(6.67% ). there is a strong correlation between the COX-2 expression and histological grades of
osteosarcoma ( P < 0. 05, P = 0. 033 ) , the positive rate of COX-2 frill increase with the histological grades of osteosarcoma. besides, there is a conelation between COX-2 and lung metastasis ( P < 0. 05, P = 0. 008 ). P-gp carries the chemical drugs out from the tumor cells by the special pump and prevents the tumor cells from being killed by the chemical therapy drugs, recent researches show that there is a negative correlation between P-gp and the sensitive of the drugs of chemical therapy and surviva
骨肿瘤是威胁人类生命健康的疾病，尤以恶性骨肿瘤为甚。在我国良性骨肿瘤占骨肿瘤及瘤样病变发病的48.45％，发病年龄多发生在11-30岁，其中，男女发病率之比约为1.56:1.00，以骨软骨瘤为最多(31.60％)，其次为骨巨细胞瘤(23.96％)。恶性肿瘤占49.30％，发病年龄也多在11-30岁，骨肉瘤的发病率占恶性骨肿瘤的76.21％，居首位。当前，对骨肿瘤的治疗方法有很多种，其中包括外科手术，放疗，化疗，免疫疗法，中草药疗法等。其中化疗在骨肿瘤的治疗中的作用越来越重要，术前化疗，手术切除，术后化疗，这一治疗模式比单纯的手术切除要有效得多。而当今造成化疗失败的主要原因是产生化疗药物的耐药，肿瘤产生耐药的机制较为复杂，目前认为，肿瘤产生多药耐药最重要的分子机制是跨膜药泵基因的扩增或过表达，这类基因以MDR1(Mutidrug resistance-1)最为重要，其表达产物为一种膜相关糖蛋白-P-糖蛋白(P-glycoprotein，P-gp)。近年来研究发现，前列腺素(PGs)和一氧化氮(NO)在肿瘤的发展和血管生成过程中起着重要的作用。环氧合酶(COX)是前列腺素合成过程中的重要限速酶，环氧合酶-2(COX-2)的过度表达通过抑制凋亡、增加细胞外基质蛋白粘附、促进肿瘤血管生成等机制参与肿瘤的发生发展。本实验的研究目的是探讨环氧合酶-2(COX-2)和P-耐药糖蛋白(P-gp)在骨肉瘤组织标本中表达的临床意义及其相关性。
材料和方法
采用免疫组织化学方法(S-P法)检测31例骨肉瘤组织蜡块标本中COX-2和P-gp蛋白的表达情况，同时另取30例骨软骨瘤组织蜡块标本作为对照组，来对比它们之间的表达是否存在差异，并探讨其意义。
结果
骨肉瘤组织中COX一和P一gP蛋白的阳性表达率分别为61 .30%和58.
10%，显著高于对照组骨软骨瘤组织6.67%和10.00%(P<0.05)。COX-2
的表达与骨肉瘤组织学分级、肺转移有关(P=0.033，0.008)。P一gP的表达
与肺转移转移有关(P二0.022)，而与组织学分级关系不大(P二0.845)。
COX一的表达与P一gP的表达密切相关(r=0.398，P二0.027)。二者的表
达与患者的性别、年龄无关。
讨论
近年莱，根据流行病学的调查统计，非街体类抗炎药(NsAIDS)确实有
降低大肠癌发病率和死亡率的作用，而NSAIDs这一作用的发挥是通过对
环氧合酶一(cox一)的抑制效应而产生的。环氧合酶是前列腺素合成过程
中的重要的限速酶，目前发现，该酶有两种亚型，即COX一l和COX一。两者
虽然结构相似，但因其生物学特性上的差异，在正常组织和病理组织中发挥
着不同的作用。尤其在正常组织中不表达或是低表达的COX一基因，一旦
受到外界因素的刺激，就会被迅速而大量的诱导表达。本组研究资料显示，
31例骨肉瘤组织中，COX一阳性表达率为61 .30%，显著高于对照组骨软
骨瘤组织6.67%(P<0.05)。COX一的表达与骨肉瘤病变组织的组织学分
级呈正相关(P<0.05，P二0.033)。随着组织学分级的增高，COXZ的阳性
表达率增加。此外，COX一的表达与肺转移有关(P<0.05，P二0.008)。P-
gP通过其特有的”药泵”作用将化疗药物泵出肿瘤细胞，从而保护肿瘤细胞
免受化疗药物的细胞毒性作用。近年来的研究表明，P一gP的表达与肿瘤的
化疗敏感性和病人的生存时间呈负相关。本组P一gP的阳性表达率是58.
10%，明显高于骨软骨瘤组织10.00%(P<0.05)，P一gP的表达与肺转移有
关(P<0.05)。cox一和P一gP参与骨肉瘤的发生和发展过程，并且它们之
间具有协同作用。选择性的应用COX一抑制剂可能会预防肿瘤耐药的发
生，并且提高肿瘤化疗的疗效。
结论
在骨肉瘤中，COX一2、P一gP参与肿瘤的发生和发展过程，并且它们之间
具有协同作用。
Objective
The tumor of bone is a dangerous diseases for human, especially maliganant bone tumors, in our country ,it is 48.45% that the benignbone tumors account for the bone tumors and tumorous pathological changes,the age ,which the disease occurs, is from 11 to 30, man/woman is about 1. 56:1. 00, the osteochon-droma is the maximum ( 31. 60% ) , the second is the giant cell tumor of bone (23.96% ). it is 49. 30% that maliganant tumors account for the bone tumors and tumorous pathological changes,the age, which the disease occurs, is from 11 to 30, the osteosarcoma is the maximum(76. 21% ). nowadays,the methods of treating the tumor of bone are very many, including operation, chemic therapeutics , radiative therapeutics, immune therapeutics, therapeutics of Chinese traditional medicine, and so on . The chemic therapeutics is more and more important for the treatment of bone tumor . the mode, ( chemic therapeutics, operation , chemic therapeutics) is much effective than operation . but the reason that caused the chemi
c therapies failure is (drug resistance . the mechanism of the drug resistance is very complicated, now most think that the important mechanism of the drug resistance is that the gene of MDR1 overexpressed,itS outcome is a glycoprotein( P-gp). recent years,the reseaches have found that PCs and NO is very important for the development and the angiogenesis of tumor. COX is a pivotal limiting enzyme during the synthetical process of PCs . COX-2 takes part in the carcinogenesis and development of osteosarcoma by inhibiting apoptosis, enhancing the adhere of extracellular matrix and accelerating the angiogenesis of tumor, the aim of the experence is to investigate the expression of P-gp and COX-2 in osteosarcoma , as well to explore their relationship and clinical significance.
Materials and Methods
COX-2 and P-gp were detected by S-P immunohistochemical method in 31 cases of paraffin-embedded sections of osteosarcoma. at the same time, I took 30 cases of paraffin-embedded sections of osteochondroma as control groups, and conformed if there was difference between the two groups and discussed the significance.
Results
The positive rate of COX-2 and P-gp in human osteosarcoma was 61. 30% and 58. 10% , respectively. Significantly higher than osteochondroma (6. 67% , 10. 00% ). The expression of COX-2 was significantly associated with the histo-logical grades , lung metastasis ( P = 0. 033, 0. 008,). The expression of P-gp was significantly correlated with lung metastasis (P =0.022) , but not with his-tological grades ( P = 0. 845 ). There was a significant correlation between the expression of COX-2 and P-gp (r =0. 398,P =0.027). Conditions such as age,sex had no relation with the expression of COX-2 or P-gp.
Discussion
recent years,bases the epidemical survey, NSAIDS can strictly reduce the incidence and mortality of colon cancer, but NSAIDS operate by inhibiting COX-2. COX is a pivotal limiting enzyme during the synthetical process of PCs. by now, we have found that it has 2 subtypes,namely COX-1 and COX-2. they have similar structure but they have different biological characteristics, they play different functions in normal tissues and pathological tissues, in normal tissues, COX-2 gene does not express or express very lower. It will largely express as soon as it is activated by somethings. In my experience, The positive rate of COX-2 was 61. 30%. Significantly higher than osteochondroma(6.67% ). there is a strong correlation between the COX-2 expression and histological grades of
osteosarcoma ( P < 0. 05, P = 0. 033 ) , the positive rate of COX-2 frill increase with the histological grades of osteosarcoma. besides, there is a conelation between COX-2 and lung metastasis ( P < 0. 05, P = 0. 008 ). P-gp carries the chemical drugs out from the tumor cells by the special pump and prevents the tumor cells from being killed by the chemical therapy drugs, recent researches show that there is a negative correlation between P-gp and the sensitive of the drugs of chemical therapy and surviva
引文
1. Ueda K, Cornwell MM, Gottesman MM, et al. The mdrl gene, responsible for multidrug resistance, codes for P-glycoprotein. Biochem Biophys Res Commun 1986; 141:956-962
2. Tanigawa N, Lu C, Mitsui T, et al. Quantitation of sinusoidlike vessels in hepatocelluar carcinoma: its clinical and prognostic significance. Hepatology 1997;26: 1216-1223
3. An FQ, Matsuda M, Fuji H, et al. Expression of vascular endothelial growth factor in surgical specimens of hepatocellular carcinoma. J Cancer Res Clin Oncol 2000; 126: 153-160
4. Chiarugi V, Magnelli L, Gallo O, et al. Cox-2, iNOS and p53 as playmarkers of tumor angiogenesis(review). Int J Mol Med 1998;2:715-719
5. Uefjii K, Ichikura T, Mochizuki H, et al. Cyclooxygenase-2 expression is related to prostaglandin biosynthesis and angiogenesis in human gastric cancer. Clin Cancer Res 2000;6: 135-138
6. Price CHG. The prognosis of osteosarcoma. Br J Radiol, 1996,39,181～188
7. Thun MJ, Namboodiri MM, Calle EE, et al. Aspirin use and risk of fatal cancer. Cancer Res, 1993,53: 1322-1327
8．郑燕，管杰，徐忠法．COX-2及其抑制剂与结直肠肿瘤．肿瘤防治杂志．2002，9(4)：487-490
9. Fujita T, Matsui M, Takaku K, et al. Size and invasion dependent increase in cyclooxygenase 2 levels in human colorectal carcinomas. Cancer Res, 1998,58:4823-4826
10. Tsujii, M, and Dubois, R. N. Alterations in cellular adhesion and apoptosis in epithelial cells overexpressing prostaglandin endoperoxide synthase 2. Cell. 1995 Nov 3;83(3):493-501
11. Gottesman MM, Pastan I, Ambudkar SV. P-glycoprotein and multidrug resistance. Curr Opin Genet Dev 1996 ;6:610-617
12. Johnstone, R. W., Ruefli A. A, and Smyth, M.J. Multiple physiological
functions for multidrug transporter P-glycoprotein? Trends Biochem Sci. 2000 Jan;25(1):1-6
13．石王伟，邹倩，黄利鸣等．宫颈癌组织中拓扑异构酶Ⅱ、P-糖蛋白的表达及其临床意义．肿瘤防治杂志．2002，9(4)：395-397
14．孙斌，王延军，王兰芳等．乳腺癌组织中P53蛋白与P-耐药糖蛋白表达水平的关系．肿瘤防治杂志．2001，8(3)：241-242
15. Ratnasinghe D, Daschner P. J, Anver M, et al. Cyclooxygenase-2, P-glycoprotein-170 and Drug Resisitance; Is Chemoprevetion Against Multidrug Resistance Possible? Anticancer Res 2001,21:2141-2148
16. Vimal A, Patel, Michael J. Dunn, and Andey Sorokin. Regulation of MDR-1 (P-glycoprotein) by Cyclooxygenase-2. J Biol Chem 2002,277: 38915-38920
17. Chambers TC, McAvoy EM, Jacobs JW, et al. Protein Kinase C phosphorylates P-glycoprotein in multidrug resistant human KB carcinoma cells. J Biol Chem 1990,265:7679-7686
18. van Helvoort A, Smith A J, Sprong H, et al. MDR1 P-glycoprotein is a lipid translocase of broad specificity, while MDR1 P-glycoprotein specifically translocates phosphatidylcholine. Cell. 1996 Nov 1;87(3):507-17
19. Moore RJ, Zweifel BS, Heuvelman DM, et al. Enhanced antitumor activity by co-administration of celecoxib and the chemotherapeutic agents cyclophosphamide and 5-FU. Proc Am Ass for Cancer Res 2000,41:409
2. Tanigawa N, Lu C, Mitsui T, et al. Quantitation of sinusoidlike vessels in hepatocelluar carcinoma: its clinical and prognostic significance. Hepatology 1997;26: 1216-1223
3. An FQ, Matsuda M, Fuji H, et al. Expression of vascular endothelial growth factor in surgical specimens of hepatocellular carcinoma. J Cancer Res Clin Oncol 2000; 126: 153-160
4. Chiarugi V, Magnelli L, Gallo O, et al. Cox-2, iNOS and p53 as playmarkers of tumor angiogenesis(review). Int J Mol Med 1998;2:715-719
5. Uefjii K, Ichikura T, Mochizuki H, et al. Cyclooxygenase-2 expression is related to prostaglandin biosynthesis and angiogenesis in human gastric cancer. Clin Cancer Res 2000;6: 135-138
6. Price CHG. The prognosis of osteosarcoma. Br J Radiol, 1996,39,181～188
7. Thun MJ, Namboodiri MM, Calle EE, et al. Aspirin use and risk of fatal cancer. Cancer Res, 1993,53: 1322-1327
8．郑燕，管杰，徐忠法．COX-2及其抑制剂与结直肠肿瘤．肿瘤防治杂志．2002，9(4)：487-490
9. Fujita T, Matsui M, Takaku K, et al. Size and invasion dependent increase in cyclooxygenase 2 levels in human colorectal carcinomas. Cancer Res, 1998,58:4823-4826
10. Tsujii, M, and Dubois, R. N. Alterations in cellular adhesion and apoptosis in epithelial cells overexpressing prostaglandin endoperoxide synthase 2. Cell. 1995 Nov 3;83(3):493-501
11. Gottesman MM, Pastan I, Ambudkar SV. P-glycoprotein and multidrug resistance. Curr Opin Genet Dev 1996 ;6:610-617
12. Johnstone, R. W., Ruefli A. A, and Smyth, M.J. Multiple physiological
functions for multidrug transporter P-glycoprotein? Trends Biochem Sci. 2000 Jan;25(1):1-6
13．石王伟，邹倩，黄利鸣等．宫颈癌组织中拓扑异构酶Ⅱ、P-糖蛋白的表达及其临床意义．肿瘤防治杂志．2002，9(4)：395-397
14．孙斌，王延军，王兰芳等．乳腺癌组织中P53蛋白与P-耐药糖蛋白表达水平的关系．肿瘤防治杂志．2001，8(3)：241-242
15. Ratnasinghe D, Daschner P. J, Anver M, et al. Cyclooxygenase-2, P-glycoprotein-170 and Drug Resisitance; Is Chemoprevetion Against Multidrug Resistance Possible? Anticancer Res 2001,21:2141-2148
16. Vimal A, Patel, Michael J. Dunn, and Andey Sorokin. Regulation of MDR-1 (P-glycoprotein) by Cyclooxygenase-2. J Biol Chem 2002,277: 38915-38920
17. Chambers TC, McAvoy EM, Jacobs JW, et al. Protein Kinase C phosphorylates P-glycoprotein in multidrug resistant human KB carcinoma cells. J Biol Chem 1990,265:7679-7686
18. van Helvoort A, Smith A J, Sprong H, et al. MDR1 P-glycoprotein is a lipid translocase of broad specificity, while MDR1 P-glycoprotein specifically translocates phosphatidylcholine. Cell. 1996 Nov 1;87(3):507-17
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