病理性瘢痕组织Ki67核增殖抗原表达与形态学观察的研究
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摘要
目的 (1)研究Ki67核增殖抗原在增生性瘢痕和瘢痕疙瘩及正常皮肤组织中的表达水平和分布特点，探讨Ki67核增殖抗原检测病理性瘢痕中细胞增殖的可行性。(2)观察病理性瘢痕的形态学变化，使用成纤维细胞平均密度计数指标，探讨Ki67核增殖抗原在病理性瘢痕组织中表达的意义；以期为临床对增生性瘢痕和瘢痕疙瘩的诊断与鉴别诊断，提供一定的依据与参考。方法 采用免疫组织化学染色方法(SP法)和常规HE染色光镜对18例增生性瘢痕、7例瘢痕疙瘩及25例自体正常皮肤，进行Ki67核增殖抗原在组织中的定位与表达量的检测、病理形态学观察与成纤维细胞平均密度计数，并进行相关的统计学分析。结果 Ki67核增殖抗原在基底层的表皮角朊细胞与真皮成纤维细胞的细胞核中出现阳性定位表达；其阳性表达量表现为：瘢痕疙瘩明显高于增生性瘢痕，增生性瘢痕明显高于正常皮肤，均存在显著性差异(P＜0.05)。形态学观察见：瘢痕疙瘩和增生性瘢痕的表皮形态异常。增生性瘢痕和瘢痕疙瘩组织中的真皮成纤维细胞数目明显多于正常皮肤，存在显著性差异(P＜0.05)；增生性瘢痕增生期亦明显多于成熟期，存在显著性差异(P＜0.05)；但成纤维细胞平均密度计数在瘢痕疙瘩与增生性瘢痕增生期之间，差异无显著性(P＞0.05)。结论 增生性瘢痕和瘢痕疙瘩组织中的基底表皮角朊细胞和成纤维细胞增殖活跃，Ki67核增殖抗原可用来检测病理性瘢痕组织中的细胞增殖状况，是一个客观性指标。Ki67核增殖抗原指标可作为增生性瘢痕和瘢痕疙瘩诊断与鉴别诊断的一个有价值的参考指标。
Objective (1) To investigate the characteristics of expression and distribution of Ki67 nuclear proliferating antigen in the hypertrophic scar (HS), keloid (K) and normal skin (NS) tissues, for identifying the possibility of Ki67 nuclear proliferating antigen detecting keratinocyte and fibroblast proliferation in the pathological scars. (2) To explore the interaction between the localization and expression characteristics of Ki67 nuclear proliferating antigen and morphological changes observed by light microscopy methods with the label of mean density of fibroblast in the HS, K, and NS tissues; and investigate if Ki67 nuclear proliferating antigen can be as a useful reference to the diagnosis and differential diagnosis of HS and K. Methods The expressive quantity and site of Ki67 nuclear proliferating antigen, the morphological change, and mean density of fibroblast were detected by using immunohistochemical staining and light microscopy methods in 18 cases of HS, 7 cases of K, and 25 cases of NS. Results (1
) The expressive site of Ki67 antigen positive was in the nucleus of basal keratinocytes and dermal flbroblasts of all HS and K tissues. The expressive quantity in K tissues was higher than HS tissues, and the expressive quantity in HS was higher than NS tissues(P< 0.05). The numbers of dermal flbroblasts in both HS and K tissues were higher than NS tissues(P<0.05).And the numbers of dermal flbroblasts of proliferating stage was higher than mature stage in HS tissues(P<0.05). However, there was not significantly difference of mean density of fibroblast between the proliferating stage of HS and K tissues(P>
0.05). (2) There was abnormal epidermis morphology in the HS and K tissues. Conclusion The fibroblast and keratinocyte proliferation in HS and K have kept a high level. Ki67 nuclear proliferating antigen may measure cell proliferation in HS and K tissues the as a valuable and objective marker, and give useful helps to the diagnosis and differential diagnosis of HS and K.
Objective (1) To investigate the characteristics of expression and distribution of Ki67 nuclear proliferating antigen in the hypertrophic scar (HS), keloid (K) and normal skin (NS) tissues, for identifying the possibility of Ki67 nuclear proliferating antigen detecting keratinocyte and fibroblast proliferation in the pathological scars. (2) To explore the interaction between the localization and expression characteristics of Ki67 nuclear proliferating antigen and morphological changes observed by light microscopy methods with the label of mean density of fibroblast in the HS, K, and NS tissues; and investigate if Ki67 nuclear proliferating antigen can be as a useful reference to the diagnosis and differential diagnosis of HS and K. Methods The expressive quantity and site of Ki67 nuclear proliferating antigen, the morphological change, and mean density of fibroblast were detected by using immunohistochemical staining and light microscopy methods in 18 cases of HS, 7 cases of K, and 25 cases of NS. Results (1
) The expressive site of Ki67 antigen positive was in the nucleus of basal keratinocytes and dermal flbroblasts of all HS and K tissues. The expressive quantity in K tissues was higher than HS tissues, and the expressive quantity in HS was higher than NS tissues(P< 0.05). The numbers of dermal flbroblasts in both HS and K tissues were higher than NS tissues(P<0.05).And the numbers of dermal flbroblasts of proliferating stage was higher than mature stage in HS tissues(P<0.05). However, there was not significantly difference of mean density of fibroblast between the proliferating stage of HS and K tissues(P>
0.05). (2) There was abnormal epidermis morphology in the HS and K tissues. Conclusion The fibroblast and keratinocyte proliferation in HS and K have kept a high level. Ki67 nuclear proliferating antigen may measure cell proliferation in HS and K tissues the as a valuable and objective marker, and give useful helps to the diagnosis and differential diagnosis of HS and K.
引文
1. Linares HA. From wound to scar. Bums, 1996;22:339-352
2. Brissett AE, Sherris DA. Scar contractures, hypertrophic scars, and keloids [J]. Facial Plast Surg,2001 ;17(4):263-272
3. Cohen IK, Diegelmann RF, Lindblad WJ. Wound healing. Biochemical clinical
aspect.philadephia wb Sanders, 1992; 500-525
4. Chiper CC, Simman R, Hatch G, et al. Mypfibroblast phenotype and apoptosis in keloid and palmar fibroblasts in vitro [J]. Cell Death Differ, 2000; 7(2): 166-176
5. Niessen FB, Andriessen MP, Schalkwijk J, et al. Keratinocyte-derived growth factors play a role in the formation of hypertrophic scars[J]. J Pathol, 2001;194(2):207-216
6. Prathiba V, Rao KS, Gupta PD. Altered expression of keratins during abnormal wound healing in human skin [J]. Cytobios, 2001; 104(405): 43-51
7. Khalid H, Shibata S, Kishikawa, et, al. Immunohistochemical analysis of progesterone receptor and Ki67 Labeling index in astorcytic tumors [J]. Cancer, 1997;80 (11):2133-2140
8. Schluter C, Duchrow M, Woblenberg C, et al. The cell proliferation associated antigen of antibody ki67: A very large ubiquitous nuclear protein with numerous repeated elements representing a new kind of cell cycle-maintaining proteins [J], J Cell Biol, 1993;23:5131
9．王允，周清华，陈桂枝，等．核增殖抗原表达基因Ki67与人肺癌临床病理生理特征及其预后的关系[J]．中国胸心血管外科临床杂志，2001；8(2)：103-106
10. Swahney N, Hall PA. Ki67—structure, function, and new antibody [J]. J Pathol, 1992;168:161
11. Toki T, Kubota J, Luxin, et al. Immuonohistochemical analysis of CA125, DA19-9, and Ki67 in stage Ⅲ or Ⅳ endonetriosis [J]. Acta Obset Gynecol Stand,2000;79:771-776
12. Mullerat J, Deroide F, Winslet MC, et al. Proliferation and p53expression in anal cancer precursor lesions [J]. Anticancer Res, 2003;23(3C): 2995-9
13. Heinz C, Hudde T, Heise K, et al. Antiproliferative effect of mycophenolate mofetil on cultured human Tenon fibroblasts [J]. Graefes Arch Clin Exp Ophthalmol, 2002; 240(5):408-14
14. Terrier-Lacombe M J, Guillou L, Maire G, et al. Dermatofibrosarcoma protuberans,
giant cell fibrosarcoma, and hybrid lesions in children: clinicopathologic comparative analysis of 28 cases with molecular data—a study from the French Federation of Cancer Centers Sarcoma Group [J]. Am J Surg Pathol, 2003; 27(1):27-39
15. Heilborn JD, Nilsson MF, Kratz G, et al. The cathelicidin anti-microbial peptide LL-37 is involved in re-epithelialization of human skin wounds and is lacking in chronic ulcer epithelium [J].J Invest Dermatol, 2003; 120(3):379-89
16. Selman M, Ruiz V, Cabrera S, et al. TIMP-1, -2, -3, and -4 in idiopathic pulmonary fibrosis. A prevailing nondegradative lung microenviroment ? [J]. Am J Physiol Lung Cell Mol Physiol, 2000;279(3):L562-74
17. Siddiqui MR, Moreira AL, Negesse Y, et al. Local nerve damage in leprosy does not lead to an impaired cellular immune response or decreased wound healing in the skin [J]. J Infect Dis, 2002; 186(2):260-5
18. Suomela S, Kariniemi AL, Impola U, et al. Matrix metalloproteimase-19 is expressed by keratinocytes in psoriasis [J].Acta Derm Venereol, 2003;83(2): 108-14
19. Teofoli P, Barduagni S, Ribuffo M, et al. Expression of Bcl-2,p53,c-jun and c-fos protooncogenes in keloids and hypertroghic scars [J].J Dermatol Sci, 1999; 22(1):31-7
20．孙同柱，付小兵，顾小曼，等．PCNA在增生性瘢痕和慢性溃疡中的表达及意义[J]．中华整形外科杂志，2002；18(4)：232-3
21．汪琴，吴宗耀．61例烧伤肥厚性瘢痕形态学观察与细胞增殖活性的研究[J]．第三军医大学学报，2001；23(7)：844-6
22. Gerdes J, Lili, Schlueter C, et al. Immunobiochemical and molecular biologic charaterization of cell proliferation—associated nuclear antigen that is defined by monoclonal antibody Ki67 [J].Am J Pathology, 1991; 138(4):867-873
23. Fonatsch C, Duchrow M,Rieder H,et al. Assignment of the human Ki67 gene (Mki67) to 10q25 qter [J].Genomics, 1991; 11(2):476-477
24. Martin P. Wound healing—aiming for perfect skin regeneration [J]. Science, 1997; 276(4):75-81
25. Desmouliere A, Redard M, Darbu L. Apoptosis mediates the decrease in cellularity during the transition between granalution tissue and scar [J].Am J Pathol, 1995; 146(1):56-66
26．付小兵，王德文主编．创伤修复基础．北京：人民军医出版社，1996：122-124
27．商庆新，袁荣，王炜．反义TGF-β1抑制瘢痕疙瘩成纤维细胞增殖的研究．中华整形外科杂志，2001；17(2)：325-27
28. Tredget EE, Nedelec B, Scott PG, et, al. Hypertrophic scars, keloids and contractures : the cellular and molecular basis therapy. Surgical Clinics of North America, 1997;77:701-730
29..Hirodin, Shen J, Yoshiharu M. Proliferating activity of dermal fibroblasts in keloids and hypertrophic scars [J].Acta Derm Venereol, 1995;75:102-4
30．黎鳌，杨宗城主编．烧伤治疗学(第二版)．北京：人民卫生出版社，1997；540-545
31. Ehrlich HP, Desmuliere A, Digeiman RF, et al. Morphological and immunochemical differences between keloids and hypertrophic scars [J].Am J Pathol, 1994; 105-13
32. Berman B, Bieley HC. Adjunct therapies to surgical management of keloids[J]. Dermatol Surg, 1996;22(1): 126-130
2. Brissett AE, Sherris DA. Scar contractures, hypertrophic scars, and keloids [J]. Facial Plast Surg,2001 ;17(4):263-272
3. Cohen IK, Diegelmann RF, Lindblad WJ. Wound healing. Biochemical clinical
aspect.philadephia wb Sanders, 1992; 500-525
4. Chiper CC, Simman R, Hatch G, et al. Mypfibroblast phenotype and apoptosis in keloid and palmar fibroblasts in vitro [J]. Cell Death Differ, 2000; 7(2): 166-176
5. Niessen FB, Andriessen MP, Schalkwijk J, et al. Keratinocyte-derived growth factors play a role in the formation of hypertrophic scars[J]. J Pathol, 2001;194(2):207-216
6. Prathiba V, Rao KS, Gupta PD. Altered expression of keratins during abnormal wound healing in human skin [J]. Cytobios, 2001; 104(405): 43-51
7. Khalid H, Shibata S, Kishikawa, et, al. Immunohistochemical analysis of progesterone receptor and Ki67 Labeling index in astorcytic tumors [J]. Cancer, 1997;80 (11):2133-2140
8. Schluter C, Duchrow M, Woblenberg C, et al. The cell proliferation associated antigen of antibody ki67: A very large ubiquitous nuclear protein with numerous repeated elements representing a new kind of cell cycle-maintaining proteins [J], J Cell Biol, 1993;23:5131
9．王允，周清华，陈桂枝，等．核增殖抗原表达基因Ki67与人肺癌临床病理生理特征及其预后的关系[J]．中国胸心血管外科临床杂志，2001；8(2)：103-106
10. Swahney N, Hall PA. Ki67—structure, function, and new antibody [J]. J Pathol, 1992;168:161
11. Toki T, Kubota J, Luxin, et al. Immuonohistochemical analysis of CA125, DA19-9, and Ki67 in stage Ⅲ or Ⅳ endonetriosis [J]. Acta Obset Gynecol Stand,2000;79:771-776
12. Mullerat J, Deroide F, Winslet MC, et al. Proliferation and p53expression in anal cancer precursor lesions [J]. Anticancer Res, 2003;23(3C): 2995-9
13. Heinz C, Hudde T, Heise K, et al. Antiproliferative effect of mycophenolate mofetil on cultured human Tenon fibroblasts [J]. Graefes Arch Clin Exp Ophthalmol, 2002; 240(5):408-14
14. Terrier-Lacombe M J, Guillou L, Maire G, et al. Dermatofibrosarcoma protuberans,
giant cell fibrosarcoma, and hybrid lesions in children: clinicopathologic comparative analysis of 28 cases with molecular data—a study from the French Federation of Cancer Centers Sarcoma Group [J]. Am J Surg Pathol, 2003; 27(1):27-39
15. Heilborn JD, Nilsson MF, Kratz G, et al. The cathelicidin anti-microbial peptide LL-37 is involved in re-epithelialization of human skin wounds and is lacking in chronic ulcer epithelium [J].J Invest Dermatol, 2003; 120(3):379-89
16. Selman M, Ruiz V, Cabrera S, et al. TIMP-1, -2, -3, and -4 in idiopathic pulmonary fibrosis. A prevailing nondegradative lung microenviroment ? [J]. Am J Physiol Lung Cell Mol Physiol, 2000;279(3):L562-74
17. Siddiqui MR, Moreira AL, Negesse Y, et al. Local nerve damage in leprosy does not lead to an impaired cellular immune response or decreased wound healing in the skin [J]. J Infect Dis, 2002; 186(2):260-5
18. Suomela S, Kariniemi AL, Impola U, et al. Matrix metalloproteimase-19 is expressed by keratinocytes in psoriasis [J].Acta Derm Venereol, 2003;83(2): 108-14
19. Teofoli P, Barduagni S, Ribuffo M, et al. Expression of Bcl-2,p53,c-jun and c-fos protooncogenes in keloids and hypertroghic scars [J].J Dermatol Sci, 1999; 22(1):31-7
20．孙同柱，付小兵，顾小曼，等．PCNA在增生性瘢痕和慢性溃疡中的表达及意义[J]．中华整形外科杂志，2002；18(4)：232-3
21．汪琴，吴宗耀．61例烧伤肥厚性瘢痕形态学观察与细胞增殖活性的研究[J]．第三军医大学学报，2001；23(7)：844-6
22. Gerdes J, Lili, Schlueter C, et al. Immunobiochemical and molecular biologic charaterization of cell proliferation—associated nuclear antigen that is defined by monoclonal antibody Ki67 [J].Am J Pathology, 1991; 138(4):867-873
23. Fonatsch C, Duchrow M,Rieder H,et al. Assignment of the human Ki67 gene (Mki67) to 10q25 qter [J].Genomics, 1991; 11(2):476-477
24. Martin P. Wound healing—aiming for perfect skin regeneration [J]. Science, 1997; 276(4):75-81
25. Desmouliere A, Redard M, Darbu L. Apoptosis mediates the decrease in cellularity during the transition between granalution tissue and scar [J].Am J Pathol, 1995; 146(1):56-66
26．付小兵，王德文主编．创伤修复基础．北京：人民军医出版社，1996：122-124
27．商庆新，袁荣，王炜．反义TGF-β1抑制瘢痕疙瘩成纤维细胞增殖的研究．中华整形外科杂志，2001；17(2)：325-27
28. Tredget EE, Nedelec B, Scott PG, et, al. Hypertrophic scars, keloids and contractures : the cellular and molecular basis therapy. Surgical Clinics of North America, 1997;77:701-730
29..Hirodin, Shen J, Yoshiharu M. Proliferating activity of dermal fibroblasts in keloids and hypertrophic scars [J].Acta Derm Venereol, 1995;75:102-4
30．黎鳌，杨宗城主编．烧伤治疗学(第二版)．北京：人民卫生出版社，1997；540-545
31. Ehrlich HP, Desmuliere A, Digeiman RF, et al. Morphological and immunochemical differences between keloids and hypertrophic scars [J].Am J Pathol, 1994; 105-13
32. Berman B, Bieley HC. Adjunct therapies to surgical management of keloids[J]. Dermatol Surg, 1996;22(1): 126-130