Smad_4蛋白和P～(21ras)蛋白的表达在胰腺癌及壶腹周围癌中的临床意义
|
摘要
背景：DPC_4基因是一个抑癌基因，K-ras基因是癌基因，两者均具有
胰腺癌特异性，它们的基因产物分别为Smad_4和P~(21ras)蛋白。目的：分析抑癌蛋白
Smad_4和癌蛋白P~(21ras)在胰腺癌和壶腹周围癌组织中的表达及其与胰腺癌的临床和
病理特征的关系，了解两种蛋白对胰腺癌与壶腹周围癌鉴别诊断的意义。方法：
采用Smad_4和P~(21ras)的单克隆抗体和免疫组织化学EnVision方法分别检测石蜡包埋
的22例胰腺癌、15例壶腹周围癌、9例胰腺癌远处淋巴结转移组织、18例良性胰
腺病变组织和3例正常胰腺组织共67例标本中的Smad_4蛋白失活和P~(21ras)蛋白表
达情况，所有被检标本均经病理诊断证实。结果：胰腺癌中，Smad_4蛋白失活率为
50.00％（11/22）；壶腹周围癌中的失活率为13.33％（2/15）；胰腺癌远处淋巴结转
移组织中失活率为44.44％（4/9）；胰腺良性病变中为16.67％（3/18），其中慢性
胰腺炎为23.08％（3/13），胰腺囊肿和胰腺囊腺瘤未发现该蛋白失活；正常胰腺中无
该蛋白失活。P~(21ras)蛋白在胰腺癌中的表达率为77.27％（17/22）；在壶腹周围癌中表
达率为26.67％（4/15）；胰腺癌远处淋巴结转移组织中的表达率为44.44％（4/9）；良
性胰腺病变中的表达率为38.89％（7/18），其中慢性胰腺炎为38.46％（5/13），胰腺囊
腺瘤为100.00％（2/2），胰腺囊肿未发现该蛋白表达；正常胰腺中未发现该蛋白表达。
胰腺癌与壶腹周围癌、胰腺良性病变间Smad_4蛋白的失活率及P~(21ras)蛋白表达率有
显著差异（p＜0.05）。胰腺癌中Smad_4蛋白的失活率和P~(21ras)蛋白表达率与患者年
龄、性别、症状、肿块大小、肿瘤部位、病理分级、临床分期和淋巴结转移无明
显相关（p＞0.05）。各病变Smad_4蛋白失活率与P~(21ras)蛋白表达率间无相关关系（p
＞O.05）。结论：用免疫组织化学方法检测Smad_4和P~(21ras)蛋白已成为了解胰腺癌中
DPC_4和K-ras基因改变的非常敏感、特异的方法。两种蛋白的检测有助于胰腺癌
与壶腹周围癌、良性胰腺病变的鉴别诊断。在K-ras基因突变这一胰腺癌发生的早
期事件之后，DPC_4基因在促进胰腺癌进一步发展中起着重要作用。
Background: DPC4 gene is an anti-oncogene, K-ras is an oncogene,
both of them are specific markers to pancreatic carcinoma, and their products are
2 ras
protein Smad4 and P Objective: To investigate the expression of anti-oncogene
product Smad4 and oncogene product P2lras in pancreatic carcinoma and periampullar
carcinoma, and the relationship of them to the clinic pathological characteratics; to
elucidate the role of the two proteins expression on the differential diagnosis of
pancreatic carcinoma and periampullar carcinoma. Methods: 67 paraffin-embedded
and pathologically proved specimens including pancreatic carcinoma (n=22),
periampullar carcinoma (n=1 5), lymph node with pancreatic carcinoma metastasis (n9),
benign disease of pancreas (n=1 8) and normal pancreas (m=3) were
immunohistochemically stained with monoclonal primary antibodies of Smad4 and
p2lras respectively by using the EnVision system. Results: Loss of Smad4 was
detected in II specimens of 22 pancreatic carcinomas (50.00%), 2 of 15
periampullar carcinomas (13.33%), 4 of 9 lymph nodes with pancreatic carcinoma
metastasis (44.44%), 3 of 18 benign diseases of pancreas (16.67%), 3 of 13 chronic
pancreatitis (23.08%) and none of 3 normal pancreata (0.00%). The positive
2lras
expression rate of P was 77.27% in pancreatic carcinomas (17/22), 26.67% in
periampullar carcinomas (4/15), 44.44% in lymph nodes with pancreatic carcinoma
metastasis (4/9), 3 8.89% in benign diseases of pancreas (7/18), 38.46% in chronic
pancreatitis (5/13) and none in normal pancreata (0/3) respectively. The loss of
Smad4 showed significant difference in pancreatic carcinoma and periampullar
carcinoma as compared to benign disease of pancreas (p <0.05) , so did the positive
expression 0fp2lraS (p <0.05) . The loss of Smad4 was not significant related to the age,
sex, symptom, tumor size, tumor site, pathological degree, clinical stages and lymph
node metastasis (p >0.05) ,. and neither was the expression of p2lras (p >0.05) . Loss
of Smad4 was not related to the expression 0fp2lras in each disease (p >0.05) ,just as
reported in other carcinomas. Conclusions: Immunohistochemical labeling for the
Smad4 and P2lras has been shown to be an extremely sensitive and specific method for
detecting the DPC4 and K-ras alterations in pancreatic carcinoma. To detect DPC4 and
K-ras gene expression could be helpful in the differential diagnosis among
pancreatic carcinoma, periampullar carcinoma and the benign diseases. The K-ras
gene occurs early in the carcinogenesis of pancreatic carcinoma, and thereafter the
loss of DPC# plays a very important role in the progress of pancreatic carcinoma.
胰腺癌特异性，它们的基因产物分别为Smad_4和P~(21ras)蛋白。目的：分析抑癌蛋白
Smad_4和癌蛋白P~(21ras)在胰腺癌和壶腹周围癌组织中的表达及其与胰腺癌的临床和
病理特征的关系，了解两种蛋白对胰腺癌与壶腹周围癌鉴别诊断的意义。方法：
采用Smad_4和P~(21ras)的单克隆抗体和免疫组织化学EnVision方法分别检测石蜡包埋
的22例胰腺癌、15例壶腹周围癌、9例胰腺癌远处淋巴结转移组织、18例良性胰
腺病变组织和3例正常胰腺组织共67例标本中的Smad_4蛋白失活和P~(21ras)蛋白表
达情况，所有被检标本均经病理诊断证实。结果：胰腺癌中，Smad_4蛋白失活率为
50.00％（11/22）；壶腹周围癌中的失活率为13.33％（2/15）；胰腺癌远处淋巴结转
移组织中失活率为44.44％（4/9）；胰腺良性病变中为16.67％（3/18），其中慢性
胰腺炎为23.08％（3/13），胰腺囊肿和胰腺囊腺瘤未发现该蛋白失活；正常胰腺中无
该蛋白失活。P~(21ras)蛋白在胰腺癌中的表达率为77.27％（17/22）；在壶腹周围癌中表
达率为26.67％（4/15）；胰腺癌远处淋巴结转移组织中的表达率为44.44％（4/9）；良
性胰腺病变中的表达率为38.89％（7/18），其中慢性胰腺炎为38.46％（5/13），胰腺囊
腺瘤为100.00％（2/2），胰腺囊肿未发现该蛋白表达；正常胰腺中未发现该蛋白表达。
胰腺癌与壶腹周围癌、胰腺良性病变间Smad_4蛋白的失活率及P~(21ras)蛋白表达率有
显著差异（p＜0.05）。胰腺癌中Smad_4蛋白的失活率和P~(21ras)蛋白表达率与患者年
龄、性别、症状、肿块大小、肿瘤部位、病理分级、临床分期和淋巴结转移无明
显相关（p＞0.05）。各病变Smad_4蛋白失活率与P~(21ras)蛋白表达率间无相关关系（p
＞O.05）。结论：用免疫组织化学方法检测Smad_4和P~(21ras)蛋白已成为了解胰腺癌中
DPC_4和K-ras基因改变的非常敏感、特异的方法。两种蛋白的检测有助于胰腺癌
与壶腹周围癌、良性胰腺病变的鉴别诊断。在K-ras基因突变这一胰腺癌发生的早
期事件之后，DPC_4基因在促进胰腺癌进一步发展中起着重要作用。
Background: DPC4 gene is an anti-oncogene, K-ras is an oncogene,
both of them are specific markers to pancreatic carcinoma, and their products are
2 ras
protein Smad4 and P Objective: To investigate the expression of anti-oncogene
product Smad4 and oncogene product P2lras in pancreatic carcinoma and periampullar
carcinoma, and the relationship of them to the clinic pathological characteratics; to
elucidate the role of the two proteins expression on the differential diagnosis of
pancreatic carcinoma and periampullar carcinoma. Methods: 67 paraffin-embedded
and pathologically proved specimens including pancreatic carcinoma (n=22),
periampullar carcinoma (n=1 5), lymph node with pancreatic carcinoma metastasis (n9),
benign disease of pancreas (n=1 8) and normal pancreas (m=3) were
immunohistochemically stained with monoclonal primary antibodies of Smad4 and
p2lras respectively by using the EnVision system. Results: Loss of Smad4 was
detected in II specimens of 22 pancreatic carcinomas (50.00%), 2 of 15
periampullar carcinomas (13.33%), 4 of 9 lymph nodes with pancreatic carcinoma
metastasis (44.44%), 3 of 18 benign diseases of pancreas (16.67%), 3 of 13 chronic
pancreatitis (23.08%) and none of 3 normal pancreata (0.00%). The positive
2lras
expression rate of P was 77.27% in pancreatic carcinomas (17/22), 26.67% in
periampullar carcinomas (4/15), 44.44% in lymph nodes with pancreatic carcinoma
metastasis (4/9), 3 8.89% in benign diseases of pancreas (7/18), 38.46% in chronic
pancreatitis (5/13) and none in normal pancreata (0/3) respectively. The loss of
Smad4 showed significant difference in pancreatic carcinoma and periampullar
carcinoma as compared to benign disease of pancreas (p <0.05) , so did the positive
expression 0fp2lraS (p <0.05) . The loss of Smad4 was not significant related to the age,
sex, symptom, tumor size, tumor site, pathological degree, clinical stages and lymph
node metastasis (p >0.05) ,. and neither was the expression of p2lras (p >0.05) . Loss
of Smad4 was not related to the expression 0fp2lras in each disease (p >0.05) ,just as
reported in other carcinomas. Conclusions: Immunohistochemical labeling for the
Smad4 and P2lras has been shown to be an extremely sensitive and specific method for
detecting the DPC4 and K-ras alterations in pancreatic carcinoma. To detect DPC4 and
K-ras gene expression could be helpful in the differential diagnosis among
pancreatic carcinoma, periampullar carcinoma and the benign diseases. The K-ras
gene occurs early in the carcinogenesis of pancreatic carcinoma, and thereafter the
loss of DPC# plays a very important role in the progress of pancreatic carcinoma.
引文
1. Silverberg E, Lubera JA. Cancer statistics. Cancer J Clin, 1989;3 :3-39.
2．王国民，赵森．胰腺癌的放射治疗进展．肿瘤，1998；18(1)：52-53．
3. Edge SB, Schmieg REJ, Rosenlof LF. Pancreas cancer resection outcome in American University in 1989-1990. Cancer, 1993;71:3502-3508.
4. Andre T, Balosso J, Louvet C, et al. Adenocarcinoma of pancreas. General characteristics. Presse Med, 1998;27(11): 533-536.
5. Almoguerra C, Shibata D, Forrester K, Martin J, et al. Most human carcinomas of the exocrine pancreas contain mutant c-K-ras genes. Cell, 1988;53:549-554.
6. Omata M, Tada M. General rules for the study of pancreatic cancer by molecular biological aspect. Nippon Geka Gakkai Zasshi, 2000; 101 (2): 233-236.
7. Banerjes SK, Makdisi WF, Weston AP, et al. A two-step enriched-nested PCR technique enhances sensitivity for detection of codon12 K-ras mutations in pancreatic adenocarcinoma. Pancreas, 1997; 15: 16-24.
8. Urban T, Ricci S, Grange JD, et al. Detection of c-K-ras mutation by PCR/RFLP analysis and diagnosis of pancreatic adenocarcinomas. J Natl Cancer Inst, 1993; 85: 2008-2012.
9. Motojima K, Tsunoda T, Kanematsu T, et al. Distinguishing pancreatic carcinoma from other periampullay carcinomas by analysis of mutations in the kirsten-ras oncogene. Ann Surg, 1991; 214(6): 657-662.
10．黄新余，钱允庆，冯昌宁，等．癌基因ras产物p21在胰腺癌、胰腺导管上皮不典型增生及胰腺内分泌肿瘤中的表达．中华实验外科杂志，1996；3(1)：27-28．
11. Hahn SA, Schutte M, Kern SE, et al. DPC_4, a candidate tumor suppressor gene at human chromosome 18q21.1. Science, 1996;271: 350-353.
12. Yingling JM, Datto MB, Wong C, et al. Tumor suppressor SMAD_4 is transforming growth factor β-inducible DNA binding protein. Mol Cell Biol, 1997;17(12):7019.
13. Sabattini E, Bisgaard K, Ascani S, et al. The EnVision++ system: a new immunohistochemical method for diagnostics and research. Critical comparison with the APAAP, ChemMate, CSA, LABC, and SABC techniques. J-Clin-Pathol. 1998 Jul; 51(7): 506-511.
14. Wilentz RE, Iacobuzio Donahue CA, Argani P, et al. Loss of expression of Dpc4 in pancreatic intraepithelial neoplasia: evidence that DPC4 inactivation occurs late in neoplastic progression. Cancer Res, 2000; 60(7): 2002-2006.
15. Wilentz RE, Su GH, Dai JL, et al. Immunohistochemical labeling for dpc4 mirrors genetic status in pancreatic adenocarcinomas: a new marker of DPC4 inactivation. Am J Pathol, 2000; 156(1): 37-43.
16．刘一平，黄翠芬．DPC4／SMAD4蛋白在细胞内信号传导中的作用．军事医学科学院院刊，1999；23(4)：309-311．
17．孙苏平．TGFβ1与恶性肿瘤治疗关系研究进展．国外医学·肿瘤学分册，1998；25(增刊)：68—71．
18. Kawabata M, Imamura T, Inoue H, et al. Intracellular signaling of the TGF-beta superfamily by Smad proteins. Ann N Y Acad Sci, 1999; 886: 73-82.
19. Kleeff J, Ishiwata T, Maruyama H, et al. The TGF-beta signaling inhibitor Smad7 enhances tumorigenicity in pancreatic cancer. Oncogene, 1999; 18(39): 5363-5372.
20. Jonson T, Gorunova L, Dawiskiba S, et al. Molecular analyses of the 15q and 18q SMAD genes in pancreatic cancer. Genes Chromosomes Cancer, 1999;24(1): 62-71.
21. Chiao P J, Hunt KK, Grau AM, et al. Tumor suppressor gene Smad4/DPC4,its downstream target genes, and regulation of cell cycle. Ann N Y Acad Sci, 1999; 880: 31-37.
22. Atfi A, Buisine M, Mazars A, et al. Induction of apoptosis by DPC4, a transcriptional factor regulated by transforming growth factor-beta through stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) signaling pathway. J Biol Chem, 1997; 272(40): 24731-24734.
23. Schutte M, Hruban RH, Hedrick L, et al. DPC4 gene in various tumor types. Cancer Res, 1996; 56(11): 2527-2530.
24. Korc M. Role of growth factors in pancreatic cancer. Surg Oncot Clin N Am, 1998; 7(1): 25-41.
25. Schwarte WI, Volpert OV, Bouck NP, et al. Smad4/DPC4-mediated tumor suppression through suppression of angiogenesis. Proc Natl Acad Sci USA. 2000; 97(17): 9624-9629.
26. Stroschein SL, Wang W, Luo K. Cooperative binding of Smad proteins to two adjacent DNA elements in the plasminogen activator inhibitor-1 promoter mediates transforming growth factor beta-induced smad-dependent transcriptional activation. J Biol Chem, 1999; 274(14): 9431-9441.
27. Schutte M. DPC4/SMAD4 gene alterations in human cancer, and their functional implications. Ann Oncol, 1999; 10 Suppl 4: 56-59.
28. Daniel SL. Molecular pathology of invasive carcinoma. Ann N Y Acad Sci, 1999;880:72-82.
29．沈魁，钟守先，张圣道主编．胰腺外科，人民卫生出版社，2000．
30. Liu F, Pouponnot C, Massague J. Dual role of the Smad4/DPC4 tumor suppressor in TGF β-inducible transcriptional complexes. Genes Dev, 1997; 11 (23): 3157-3167.
31．何三光，宋茂民，沈魁．胰腺癌的基因诊断及基因治疗的研究现状．实用肿瘤学杂志，1999；13(4)：309-311．
32. Iacobuzio Donahue CA, Klimstra DS, Adsay NV, et al.Dpc-4 protein is expressed in virtually all human intraductal papillary mucinous neoplasms of the pancreas: comparison with conventional ductal adenocarcinomas. Am J Pathol 2000; 157(3): 755-761.
2．王国民，赵森．胰腺癌的放射治疗进展．肿瘤，1998；18(1)：52-53．
3. Edge SB, Schmieg REJ, Rosenlof LF. Pancreas cancer resection outcome in American University in 1989-1990. Cancer, 1993;71:3502-3508.
4. Andre T, Balosso J, Louvet C, et al. Adenocarcinoma of pancreas. General characteristics. Presse Med, 1998;27(11): 533-536.
5. Almoguerra C, Shibata D, Forrester K, Martin J, et al. Most human carcinomas of the exocrine pancreas contain mutant c-K-ras genes. Cell, 1988;53:549-554.
6. Omata M, Tada M. General rules for the study of pancreatic cancer by molecular biological aspect. Nippon Geka Gakkai Zasshi, 2000; 101 (2): 233-236.
7. Banerjes SK, Makdisi WF, Weston AP, et al. A two-step enriched-nested PCR technique enhances sensitivity for detection of codon12 K-ras mutations in pancreatic adenocarcinoma. Pancreas, 1997; 15: 16-24.
8. Urban T, Ricci S, Grange JD, et al. Detection of c-K-ras mutation by PCR/RFLP analysis and diagnosis of pancreatic adenocarcinomas. J Natl Cancer Inst, 1993; 85: 2008-2012.
9. Motojima K, Tsunoda T, Kanematsu T, et al. Distinguishing pancreatic carcinoma from other periampullay carcinomas by analysis of mutations in the kirsten-ras oncogene. Ann Surg, 1991; 214(6): 657-662.
10．黄新余，钱允庆，冯昌宁，等．癌基因ras产物p21在胰腺癌、胰腺导管上皮不典型增生及胰腺内分泌肿瘤中的表达．中华实验外科杂志，1996；3(1)：27-28．
11. Hahn SA, Schutte M, Kern SE, et al. DPC_4, a candidate tumor suppressor gene at human chromosome 18q21.1. Science, 1996;271: 350-353.
12. Yingling JM, Datto MB, Wong C, et al. Tumor suppressor SMAD_4 is transforming growth factor β-inducible DNA binding protein. Mol Cell Biol, 1997;17(12):7019.
13. Sabattini E, Bisgaard K, Ascani S, et al. The EnVision++ system: a new immunohistochemical method for diagnostics and research. Critical comparison with the APAAP, ChemMate, CSA, LABC, and SABC techniques. J-Clin-Pathol. 1998 Jul; 51(7): 506-511.
14. Wilentz RE, Iacobuzio Donahue CA, Argani P, et al. Loss of expression of Dpc4 in pancreatic intraepithelial neoplasia: evidence that DPC4 inactivation occurs late in neoplastic progression. Cancer Res, 2000; 60(7): 2002-2006.
15. Wilentz RE, Su GH, Dai JL, et al. Immunohistochemical labeling for dpc4 mirrors genetic status in pancreatic adenocarcinomas: a new marker of DPC4 inactivation. Am J Pathol, 2000; 156(1): 37-43.
16．刘一平，黄翠芬．DPC4／SMAD4蛋白在细胞内信号传导中的作用．军事医学科学院院刊，1999；23(4)：309-311．
17．孙苏平．TGFβ1与恶性肿瘤治疗关系研究进展．国外医学·肿瘤学分册，1998；25(增刊)：68—71．
18. Kawabata M, Imamura T, Inoue H, et al. Intracellular signaling of the TGF-beta superfamily by Smad proteins. Ann N Y Acad Sci, 1999; 886: 73-82.
19. Kleeff J, Ishiwata T, Maruyama H, et al. The TGF-beta signaling inhibitor Smad7 enhances tumorigenicity in pancreatic cancer. Oncogene, 1999; 18(39): 5363-5372.
20. Jonson T, Gorunova L, Dawiskiba S, et al. Molecular analyses of the 15q and 18q SMAD genes in pancreatic cancer. Genes Chromosomes Cancer, 1999;24(1): 62-71.
21. Chiao P J, Hunt KK, Grau AM, et al. Tumor suppressor gene Smad4/DPC4,its downstream target genes, and regulation of cell cycle. Ann N Y Acad Sci, 1999; 880: 31-37.
22. Atfi A, Buisine M, Mazars A, et al. Induction of apoptosis by DPC4, a transcriptional factor regulated by transforming growth factor-beta through stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) signaling pathway. J Biol Chem, 1997; 272(40): 24731-24734.
23. Schutte M, Hruban RH, Hedrick L, et al. DPC4 gene in various tumor types. Cancer Res, 1996; 56(11): 2527-2530.
24. Korc M. Role of growth factors in pancreatic cancer. Surg Oncot Clin N Am, 1998; 7(1): 25-41.
25. Schwarte WI, Volpert OV, Bouck NP, et al. Smad4/DPC4-mediated tumor suppression through suppression of angiogenesis. Proc Natl Acad Sci USA. 2000; 97(17): 9624-9629.
26. Stroschein SL, Wang W, Luo K. Cooperative binding of Smad proteins to two adjacent DNA elements in the plasminogen activator inhibitor-1 promoter mediates transforming growth factor beta-induced smad-dependent transcriptional activation. J Biol Chem, 1999; 274(14): 9431-9441.
27. Schutte M. DPC4/SMAD4 gene alterations in human cancer, and their functional implications. Ann Oncol, 1999; 10 Suppl 4: 56-59.
28. Daniel SL. Molecular pathology of invasive carcinoma. Ann N Y Acad Sci, 1999;880:72-82.
29．沈魁，钟守先，张圣道主编．胰腺外科，人民卫生出版社，2000．
30. Liu F, Pouponnot C, Massague J. Dual role of the Smad4/DPC4 tumor suppressor in TGF β-inducible transcriptional complexes. Genes Dev, 1997; 11 (23): 3157-3167.
31．何三光，宋茂民，沈魁．胰腺癌的基因诊断及基因治疗的研究现状．实用肿瘤学杂志，1999；13(4)：309-311．
32. Iacobuzio Donahue CA, Klimstra DS, Adsay NV, et al.Dpc-4 protein is expressed in virtually all human intraductal papillary mucinous neoplasms of the pancreas: comparison with conventional ductal adenocarcinomas. Am J Pathol 2000; 157(3): 755-761.