摘要
目的 研究小涎腺粘液表皮样癌(MEC)组织学分级、检测血管内皮生长因子(VEGF)的表达，计数CD_(34)标记的肿瘤内微血管密度(MVD)并分析它们间的关系及其与肿瘤临床分期和复发的关系。
材料与方法 收集1970—2002年武汉大学口腔医院经治并经病理确诊的小涎腺粘液表皮样癌121例，其中77例符合本次实验病例选择标准。复习临床资料，重新读片，采用Auclair提出的量化分级标准对本组病例行组织学分级。对其中47例有完整随访资料的病例行免疫组织化学S-P法，检测VEGF的表达、计数CD_(34)标记的肿瘤内微血管密度，并行统计分析。
结果 77例中Ⅰ级32例占41.6％，Ⅱ级24例占31.2％，Ⅲ级21例占27.2％。经x~2检验临床分期与组织学分级有关(x~2=12.065，P＜0.05)。临床分期、组织学分级与肿瘤复发均显著相关(P＜0.05，log-rank检验)。47例小涎腺MEC中MVD为27.73±8.72(15～56)／×200，MVD与组织学分级、临床分期及复发均显著相关(P＜0.05)。47例中37例VEGF染色阳性(78.7％)。不同组织学分级、临床分期间VEGF阳性表达率差异无显著性(P＞0.05)，VEGF表达与肿瘤复发无明显相关(P＞0.05，log-rank检验)。VEGF阳性组MVD(30.85±8.42)明显高于阴性组(20.71±3.73)，差异有显著性(t=5.655，P＜0.05)，VEGF表达与MVD呈正相关(r_s=0.6，P＜0.05，Spearman)。
结论 应用Auclair提出的量化分级标准对小涎腺粘液表皮样癌行组织学分级有助于判断肿瘤的生物学行为。小涎腺粘液表皮样癌中肿瘤细胞能产生VEGF促进血管生成，计数以CD_(34)标记的肿瘤内微血管密度在小涎腺MEC病理学分级诊断中及对判断肿瘤的生物学行为有一定应用价值。
Objective This study evaluated the prognostic importance of a quantitative grading system, the expression of vascular endothelial growth factor(VEGF) and intratumoral microvessel density (MVD , labeled by anti-human CD34 monoclonal antibody) in mucoepidermoid carcinoma(MEC) of intraoral salivary glands.
Materials & Methods We have clinicopathologically reviewed 77 cases of MEC originating in intraoral salivary glands that were treated at Hospital of Stomatology of Wuhan University between 1970 and 2002. Following the methods of Auclair, a quantitative grading system was used. The expression of VEGF, intratumoral microvessel density (MVD, labeled by anti-human CD34 monoclonal antibody) in 47 cases with complete follow-up materials were examined immunohistochemically.
Results The percentages were 41.6% for low-grade, 31.2% for intermediate-grade and 27.2% for high-grade tumors. The correlation between the TNM classification and the histologic grading was significant ( x 2=12.065, P<0.05). Both the TNM classification and the histologic grading were associated with an increased incidence of recurrence (P<0.05,log-rank test). MVD in 47 cases varied from 15~56 (27.73?.72) /x200. MVD was correlated with the histologic grading, the TNM classification and tumor recurrence(P<0.05). Positive staining for VEGF was obtained in 37 of the 47 cases (78.7%). No significant difference in VEGF expression was found among the groups. No correlation was noted between VEGF expression and recurrence(P>0.05, log-rank test). MVD was significantly higher in VGEF positive than in VGEF negative cases (30.85?.42, 20.71?.73, respectively; t=5.655, P<0.05). The correlation between the expression of VEGF and MVD was significant(rs=0.6, P<0.05, Spearman).
Conclusions Our study confirms that in MEC of intraoral salivary glands, tumor grade subdividing cases into low , intermediate and high grade by using the criteria delineated by Auclair, correlates well with prognosis. In MEC of intraoral salivary glands, tumor cells can produce VEGF to stimulate angiogenesis. MVD, labeled by anti-human CD34 monoclonal antibody, might be useful in evaluating the histopathologic grading and prognosis of patients with MEC of intraoral salivary glands.
引文
1. Stewart FW, Foote FW, Becker WF. Mucoepidermoid tumors of salivary glands. Ann Surg, 1945,122:820-844.
2. Aulclair PL, Ellis GL. Mucoepidermoid carcinoma. Ellis GL, Aulclair PL, Gnepp DR, editors. Surgical Pathology of the Salivary Glands. Philadelphia:W.B.Saunders,1991:269-298.
3. Hassanin MB, Ghosh L, Das AK, et al. Immunohistochemical and fluorescent microscopic
study of histogenesis of salivary mucoepidermoid carcinoma. J Oral Pathol Med, 1989,18(5): 291-298.
4. Chaudhry AP, Cutler LS, Leifer C, et al. Ultrastructural study of the histogenesis of salivary gland mucoepidermoid carcinoma. J Oral Pathol Med, 1989,18(7): 400-409.
5. Yin HF, Okada N, Takagi M. Apoptosis and apoptotic-related factors in mucoepidermoid carcinoma of the oral minor salivary glands. Pathol Int, 2000,50(8): 603-609.
6. Frankenthaler RA, el-Naggar AK, Ordonez NG, et al. High correlation with survival of proliferating cell nuclear antigen expression in mucoepidermoid carcinoma of the parotid gland. Otolaryngol Head Neck Surg, 1994,111(4): 460-466.
7. Fujita S, Takahashi H, Okabe H. Nucleolar organizer regions in malignant salivary gland tumors. Acta Pathol Jpn, 1992,42(I0): 727-733.
8. Suzuki M, Ichimiya I, Matsushita F, et al. Histological features and prognosis of patients with mucoepidermoid carcinoma of the parotid gland. J Laryngol Otol, 1998,112(10): 944-947.
9. Foote FW, Frazell EL. Tumors of the salivary glands. Cancer, 1953,6(6):1065-1133.
10. Auclair PL, Goode RK, Ellis GL. Mucoepidermoid carcinoma of intraoral salivary glands. Evaluation and application of grading criteria in 143 cases. Cancer, 1992,69(8):2021-2030.
11. Goode RK, Auclair PL, Ellis GL. Mucoepidermoid carcinoma of the major salivary glands: clinical and histopathologic analysis of 234 cases with evaluation of grading criteria. Cancer,1998,82(7): 1217-1224.
12. Spiro RH, Huvos AG, Berk R, et al. Mucoepidermoid carcinoma of salivary gland origin. A clinicopathologic study of 367 cases. Am J Surg, 1978,136(4): 461-468.
13. Guzzo M, Andreola S, Sirizzotti G, et al. Mucoepidermoid carcinoma of the salivary glands: clinicopathologic review of 108 patients treated at the National Cancer Institute of Milan. Ann Surg Oncol,2002, 9(7):688-695.
14. Folkman J. Tumor angiogenesis: therapeutic implications. N Engl J Med, 1971, 285(21): 1182-1186.
15. Maisonpierre PC, Suri C, Jones PF, et al. Angiopoietin-2, a natural antagonist for Tie2 that
disrupts in vivo angiogenesis. Science, 1997,277(5322): 55-60.
16. Fujieda S, Sunaga H, Tsuzuki H, et al. Expression of platelet-derived endothelial cell growth factor in oral and oropharyngeal carcinoma. Clin Cancer Res, 1998,4(7): 1583-1590.
17. Eisma RJ, Spiro JD, Kreutzer DL. Vascular endothelial growth factor expression in head and neck squamous cell carcinoma. Am J Surg, 1997,174(5): 513-517.
18. Denhart BC, Guidi AJ, Tognazzi K, et al. Vascular permeability factor/vascular endothelial growth factor and its receptors in oral and laryngeal squamous cell carcinoma and dysplasia. Lab Invest, 1997,77(6): 659-664.
19. Ishibashi H, Shiratuchi T, Nakagawa K, et al. Hypoxia-induced angiogenesis of cultured human salivary gland carcinoma cells enhances vascular endothelial growth factor production and basic fibroblast growth factor release. Oral Oncol, 2001,37(1): 77-83.
20. Doi R, Kuratate I, Okamoto E, et al. Expression of p53 oncoprotein increases intratumoral microvessel formation in human salivary gland carcinomas. J Oral Pathol Med, 1999,28(6): 259-263.
21．欧阳喈．粘液表皮样癌．见：于世凤，主编．口腔组织病理学．第4版．北京：人民卫生出版社，2000：250-253．
22. Thackray AC, et al. Histological typing of salivary gland tumors. In: International histological classification of tumors.No7.Geneva: World Health Organization,1972:23.
23．刘瑷如，刘桢，罗汉兰，等．241例涎腺粘液表皮样癌的临床病理分析，中华医学杂志，1986，66(7)：392-394．
24. Seifert G. World Health Organization Classification of tumors: Histological typing of salivary gland tumors. 2nd ed. Berlin: Springerverlag, 1991:20-21.
25. Brandwein MS, Ivanov K, Wallace DI, et al. Mucoepidermoid carcinoma: a clinicopathologic study of 80 patients with special reference to histological grading. Am J Surg Pathol,2001,25(7):835-845.
26. Jakobsson PA, Blanck C, Eneroth CM. Mucoepidermoid carcinoma of the parotid gland. Cancer, 1968,22(1): 111-124.
27. Thorvaldsson SE, Beahrs OH, Woolner LB, et al. Mucoepidermoid tumors of the major
salivary glands. Am J Surg, 1970,120(4): 432-438.
28. Accetta PA, Gray GF Jr, Hunter RM, et al. Mucoepidermoid carcinoma of salivary glands. Arch Pathol Lab Med, 1984,108(4): 321-325.
29. Nascimento AG, Amaral LP, Prado LA, et al. Mucoepidermoid carcinoma of salivary glands: a clinicopathologic study of 46 cases. Head Neck Surg, 1986,8(6): 409-417.
30. Evans HL. Mucoepidermoid carcinoma of salivary glands: a study of 69 cases with special attention to histologic grading. Am J Clin Pathol, 1984, 81(6): 696-701.
31. Dougall WC, Qian X, Peterson NC, et al. The neu-oncogene: signal transduction pathways, transformation mechanisms and evolving therapies. Oncogene, 1994,9(8): 2109-2123.
32. Batsakis JG. Staging of salivary gland neoplasms: role of histopathologic and molecular factors. Am J Surg, 1994,168(5): 386-390.
33. Press MF, Pike MC, Hung G, et al. Amplification and overexpression of HER-2/neu in carcinomas of the salivary gland: correlation with poor prognosis. Cancer Res, 1994,54(21): 5675-5682.
34．朱明华，p53基因．见：曾益新，主编．肿瘤学，第一版．北京：人民卫生出版社，1999：59-61．
35. Kiyoshima T, Shima K, Kobayashi I, et al. Expression of p53 tumor suppressor gene in adenoid cystic and mucoepidermoid carcinomas of the salivary glands. Oral Oncol, 2001,37(3): 315-322.
36. Karja VJ, Syrjanen KJ, Kurvinen AK, et al. Expression and mutations of p53 in salivary gland tumours. J Oral Pathol Med. 1997,26(5): 217-223.
37．龚建平，细胞周期与肿瘤．见：曾益新，主编．肿瘤学，第一版．北京：人民卫生出版社，1999：97-117．
38. Tubiana M, Pejovic MH, Koscielny S, et al. Growth rate, kinetics of tumor cell proliferation and long-term outcome in human breast cancer. Int J Cancer, 1989,44(1): 17-22.
39. Silvestrini R, Daidone MG, Bolis G, et al. Cell kinetics: a prognostic marker in epithelial ovarian cancer. Gynecol Oncol, 1989,35(1): 15-19.
40. Sahin AA, Ro J, Ro JY, et al. Ki-67 immunostaining in node-negative stage Ⅰ and Ⅱ breast
carcinoma:Significant correlation with prognosis. Cancer, 1991,68(3): 549-557.
41. Skalova A, Lehtonen H, von Boguslawsky K, et al. Prognostic significance of cell proliferation in mucoepidermoid carcinomas of the salivary gland: clinicopathological study using MIB 1 antibody in paraffin sections. Hum Pathol, 1994,25(9): 929-935.
42. Hicks J, Flaitz C. Mucoepidermoid carcinoma of salivary glands in children and adolescents: assessment of proliferation markers. Oral Oncol, 2000,36(5): 454-460.
43. Zhu Q, White FH, Tipoe GL. The assessment of proliferating cell nuclear antigen (PCNA) immunostaining in human benign and malignant epithelial lesions of the parotid gland. Oral Oncol, 1997,33(1): 29-35.
44. Cardoso WP, Denardin OV, Rapoport A, et al. Proliferating cell nuclear antigen expression in mucoepidermoid carcinoma of salivary glands. Sao Paulo Med J, 2000,118(3): 69-74.
45．陈坤，高岩，庞淑珍，等．增殖细胞核抗原和抑癌基因P53在粘液表皮样癌中的表达．中华口腔医学杂志，1996，31(5)：285-287．
46. Ofner D, Riedmann B, Maier H, et al. Standardized staining and analysis of argyrophilic nucleolar organizer region associated proteins (AgNORs) in radically resected colorectal adenocarcinoma--correlation with tumour stage and long-term survival. J Pathol, 1995, 175(4):441-448.
47. Ploton D, Menager M, Jeannesson P, et al. Improvement in the staining and in the visualization of the argyrophilic proteins of the nucleolar organizer region at the optical level. Histochem J, 1986,18(1):5-14.
48．王庆，汪说之，苏倩倩．涎腺粘液表皮样癌中的核仁组织导体区研究．华西口腔医学杂志，1992，10(1)：29-31．
49. Okabe M, Inagaki H, Murase T, et al. Prognostic significance of p27 and Ki-67 expression in mucoepidermoid carcinoma of the intraoral minor salivary gland. Mod Pathol, 2001, 14(10): 1008-1014.
50. Shih IM, Nesbit M, Herlyn M, et al. A new Mel-CAM (CD146)-specific monoclonal antibody, MN-4, on paraffin-embedded tissue. Mod Pathol, 1998,11(11): 1098-1106.
51. Pires FR, Shih IeM, da Cruz Perez DE, et al. Mel-CAM (CD146) expression in parotid
mucoepidermoid carcinoma. Oral Oncol, 2003,39(3): 277-281.
52．马丁，肿瘤转移的分子生物学基础，见：曾益新，主编．肿瘤学，第一版．北京：人民卫生出版社，1999：184-191．
53. Shieh YS, Chang LC, Chiu KC, et al. Cadherin and catenin expression in mucoepidermoid carcinoma: correlation with histopathologic grade, clinical stage, and patient outcome. J Oral Pathol Med, 2003,32(5): 297-304.
54．宋琦、汪说之，苏倩倩．涎腺粘液表皮样癌癌胚抗原的免疫组化研究．华西口腔医学杂志，1992，10(2)：90-92．
55．杨连甲，张海霞，孙庆妹．凝集素在正常涎腺和粘液表皮样癌中的细胞定位．中华口腔医学杂志，1990，25(2)：96-98．
56．刁桂香，胡永升，张学斌，等．涎腺肿瘤中雌、孕激素受体的表达及意义．中国肿瘤临床，1995，22(6)：421-424．
57. Sobin LH, Wittekind CH. UICC:TNM classification of maglignant tumors. 5th ed. New York:Wiley-liss, 1997:17-46
58. Mattern J, Koomagi R, Volm M. Association of vascular endothelial growth factor expression with intratumoral microvessel density and tumor cell proliferation in human epidermoid lung carcinoma. Br J Cancer, 1996,73(7):931-934
59. Tenderenda M, Rutkowski P, Jesionek-Kupnicka D, et al. Expression of CD34 in gastric cancer and its correlation with histology, stage, proliferation activity, p53 expression and apoptotic index. Pathol Oncol Res, 2001,7(2):129-134
60. Saeki T, Tanada M, Takashima S, et al. Correlation between expression of platelet-derived endothelial cell growth factor (thymidine phosphorylase) and microvessel density in early-stage human colon carcinomas. Jpn J Clin Oncol, 1997,27(4):227-230.
61. Bettencourt MC, Bauer JJ, Sesterhenn IA, et al. CD34 immunohistochemical assessment of angiogenesis as a prognostic marker for prostate cancer recurrence after radical prostatectomy. J Urol,1998,160(2):459-465
62. Obermair A, Tempfer C, Wasicky R, et al. Prognostic significance of tumor angiogenesis in endometrial cancer. Obstet Gynecol, 1999,93(3):367-371.
63. Choi HJ, Hyun MS, Jung GJ, et al. Tumor angiogenesis as a prognostic predictor in colorectal carcinoma with special reference to mode of metastasis and recurrence. Oncology 1998,55(6):575-581.
64. Tomisaki S, Ohno S, Ichiyoshi Y, et al. Microvessel quantification and its possible relation with liver metastasis in colorectal cancer. Cancer, 1996,77(8 Suppl): 1722-1728.
65. Heimburg S, Oehler MK, Papadopoulos T, et al. Prognostic relevance of the endothelial marker CD 34 in ovarian cancer. Anticancer Res,1999,19(4A):2527-2529.
66. Abdalla SA, Behzad F, Bsharah S, et al. Prognostic relevance of microvessel density in colorectal tumours. Oncol Rep, 1999,6(4):839-842.
67. Neufeld G, Cohen T, Gengrinovitch S, et al. Vascular endothelial growth factor (VEGF) and its receptors.FASEB J,1999,13(1):9-22
68．高文信，王雷，马宁．口腔粘膜癌前病变及鳞状细胞癌中VEGF的表达及意义．口腔医学研究，2002，18(5)：319-321
69. Brown LF, Berse B, Jackman RW, et al. Expression of vascular permeability factor (vascular endothelial growth factor) and its receptors in adenocarcinomas of the gastrointestinal tract. Cancer Res, 1993,53(19):4727-4735.
70. Maeda K, Chung YS, Ogawa Y, et al. Prognostic value of vascular endothelial growth factor expression in gastric carcinoma. Cancer, 1996,77(5):858-863.
71．李正江，唐平章，徐震刚，等．微血管密度和血管内皮生长因子在涎腺腺样囊性癌中的表达．中华口腔医学杂志，2001，36(3)：212-214．
72. Pammer J, Weninger W, Mildner M, et al. Vascular endothelial growth factor is constitutively expressed in normal human salivary glands and is secreted in the saliva of healthy individuals. J Pathol, 1998,186(2): 186-191.