外阴营养不良中Caspase-3，Ki-67的表达与其病变相关性的病理学研究
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摘要
目的：检测凋亡关键蛋白酶caspase-3与增殖细胞核抗原ki-67在外阴营养不良和外阴鳞癌中的表达，旨在揭示caspase-3、ki-67是否参与了外阴营养不良的病变过程。
方法：应用免疫组织化学链霉素抗生物素蛋白过氧化酶法(SP法)，检测caspase-3、ki-67蛋白在外阴营养不良、外阴鳞癌和正常外阴皮肤中的表达。
结果：1．69例外阴营养不良按照年龄分组，增生型22例、硬化苔癣型20例、混合型8例、伴不典型增生19例，其中增生型病例中绝经期前所占例数比绝经期后多，硬化苔藓型、混合型和不典型增生组病例中绝经期后所占例数分别比绝经期前多。(P=0.000)
2．①正常外阴皮肤组caspase-3蛋白表达阳性率(50％)低于硬化苔藓型外阴营养不良组(90％)(P=0.026)；硬化苔藓型营养不良组caspase-3蛋白表达阳性率高于增生型外阴营养不良组(59％)(P=0.023)；不典型增生组、外阴鳞癌组caspase-3蛋白表达阳性率分别与正常皮肤组及各型营养不良相比均显著降低(P＜0.05)。②增生型外阴营养不良组ki-67的蛋白表达阳性率(91％)高于硬化苔藓型营养不良组(60％)(P=0.019)；不典型增生组ki-67的蛋白表达阳性率(89％)高于硬化苔藓型营养不良组(P=0.019)。
结论：1．外阴营养不良的发生与性激素水平相关。
2．外阴营养不良病变的形成与表皮细胞凋亡和增殖平衡紊乱有关。
OBJECTIVE: The expressions of caspase-3 and ki-67 are tested to evaluate its effects on vulvar dystrophy and squamous cell carcinoma of vulva.
METHODS: The immunohistochemistry(SP method) is used to determine the expressions of caspase-3 and ki-67 in vulvar dystrophy, atypical hyperplasia, squamous cell carcinoma of vulva and normal vulval skin.
RESULTS: 1. According to different ages, 69 cases of vulvar dystrophies are divided into 4 groups:22 cases were diagnosed Squamous cell hyperplasia, 20 cases were diagnosed Lichen sclerosus, 8 cases were diagnosed mixed type and the rest cases were diagnosed atypical hyperplasia. Among the cases of Squamous cell hyperplasia, the cases of before menostasia were more than those of after menostasia. In the other three groups, the cases of after menostasia were more than those of before menostasia, respectively. 2. (1)The protein expression level of caspase-3 in normal vulval skin(50%) was significantly lower than that in Lichen sclerosus (90%). The protein expression level of caspase-3 in Lichen sclerosus was significantly higher than in Squamous cell hyperplasia(59%).The positive expression of caspase-3 in atypical hyperplasia and vulvar cancer were significantly lower than in normal vulval skin and vulvar dystrophy respectively.(2)The protein expression of
方法：应用免疫组织化学链霉素抗生物素蛋白过氧化酶法(SP法)，检测caspase-3、ki-67蛋白在外阴营养不良、外阴鳞癌和正常外阴皮肤中的表达。
结果：1．69例外阴营养不良按照年龄分组，增生型22例、硬化苔癣型20例、混合型8例、伴不典型增生19例，其中增生型病例中绝经期前所占例数比绝经期后多，硬化苔藓型、混合型和不典型增生组病例中绝经期后所占例数分别比绝经期前多。(P=0.000)
2．①正常外阴皮肤组caspase-3蛋白表达阳性率(50％)低于硬化苔藓型外阴营养不良组(90％)(P=0.026)；硬化苔藓型营养不良组caspase-3蛋白表达阳性率高于增生型外阴营养不良组(59％)(P=0.023)；不典型增生组、外阴鳞癌组caspase-3蛋白表达阳性率分别与正常皮肤组及各型营养不良相比均显著降低(P＜0.05)。②增生型外阴营养不良组ki-67的蛋白表达阳性率(91％)高于硬化苔藓型营养不良组(60％)(P=0.019)；不典型增生组ki-67的蛋白表达阳性率(89％)高于硬化苔藓型营养不良组(P=0.019)。
结论：1．外阴营养不良的发生与性激素水平相关。
2．外阴营养不良病变的形成与表皮细胞凋亡和增殖平衡紊乱有关。
OBJECTIVE: The expressions of caspase-3 and ki-67 are tested to evaluate its effects on vulvar dystrophy and squamous cell carcinoma of vulva.
METHODS: The immunohistochemistry(SP method) is used to determine the expressions of caspase-3 and ki-67 in vulvar dystrophy, atypical hyperplasia, squamous cell carcinoma of vulva and normal vulval skin.
RESULTS: 1. According to different ages, 69 cases of vulvar dystrophies are divided into 4 groups:22 cases were diagnosed Squamous cell hyperplasia, 20 cases were diagnosed Lichen sclerosus, 8 cases were diagnosed mixed type and the rest cases were diagnosed atypical hyperplasia. Among the cases of Squamous cell hyperplasia, the cases of before menostasia were more than those of after menostasia. In the other three groups, the cases of after menostasia were more than those of before menostasia, respectively. 2. (1)The protein expression level of caspase-3 in normal vulval skin(50%) was significantly lower than that in Lichen sclerosus (90%). The protein expression level of caspase-3 in Lichen sclerosus was significantly higher than in Squamous cell hyperplasia(59%).The positive expression of caspase-3 in atypical hyperplasia and vulvar cancer were significantly lower than in normal vulval skin and vulvar dystrophy respectively.(2)The protein expression of
引文
1. Shirer JA, Jr Ray MC. Familial occurrence of lichen sclerosuset atrophicas Case reports of a mother and daughter. Arch Dermatol, 1987, 123(4): 485-488.
2. Meyers C, Bromberg-White JL, Zhang J , et al. Infectious virions produced from a human papillomavirus type 18/16 genomic DNA chimera J. Virol, 2002, 76(10): 4723-4733.
3．张莉，金晓明，迟月明等．人乳头状瘤病毒感染与外阴营养不良相关性的研究．电子显微学报，2003，22(4)：267-270．
4．尹菊，刘立峰，周仲华．谷光甘肽、SOD软膏红外线照射治疗外阴营养不良的疗效分析．中国医科大学学报，1998，27(6)：643-644．
5．唱捷，何文翠，张建国等．人表皮生长因子与外阴白色病变的关系．中华妇产科杂志，1995，30(4)：209-211．
6．乔丽雅，李亚里，关铮．表皮生长因子受体在外阴营养不良和外阴鳞癌中的表达及其临床意义．解放军医学杂志，2000，25(2)：138-139．
7．杨树君，陈铭，陈诵芬等．外阴营养不良中fas／apol-1表达的生物学意义．中国实验诊断学．2000，4(3)：111-112．
8. Bccagli L, Cadore L. Sjogren's syndrome and vulvar cancer. Clin Exp obstet Gynecol, 1987, 14(1): 69-71.
9. Coarlson JA, Grabowski R, Chichester P, et al. Comparartive immunophenotypic study of lichen sclerosus: epidermotropicCD57~+ lymphocytes are numerous-implications for pathogenesis, Am J Dermatopathol, 2000, 22(1): 7-16.
10．勾美图，瑞华，付君等．胎盘多肽治疗外阴营养不良94例疗效分析．中国实用妇科与产科杂志，1996，12(6)：369-370．
11. Oltvai ZN, Milliman Cl, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a c onserved homolog, Bax, that accelerates programmed cell death. Cell, 1993, 74(4): 609-619.
12．欧阳玲，张淑兰，王小松等．Bcl-2、bax、P53蛋白表达与老年妇女外阴营养不良细??胞凋亡的相关性研究．中国现代医学杂志，2003，13(12)：48-50．
13. Scheistroen M, Trope C, Pettersen EO, et al. P53 protein expression in squamosu cell carcinoma of the vulva. Cancer, 1999, 85(5): 1133-1138.
14．张式暖，王亚利，王艳华等．食管癌及癌前病变组织中癌基因的表达．中国现代医学杂志，2000，10(9)：8-9．
15．李远晃，徐志林．胃癌组织中CyclinE和P53蛋白表达的研究．中国现代医学杂志，2002，12(1)：23-25．
16．宁玉梅，石一复．PCNA在外阴癌及癌前病变中的表达及其临床意义[J]．实用肿瘤杂志，2000，15(4)：234-236．
17．欧阳玲，马晓欣，张淑兰．Bcl-2、Bax蛋白表达与外阴营养不良细胞凋亡的相关性研究[J]．中国医科大学学报 2000，29(6)：436-437．
18. Schuler M, Bossy-Wetzel E, Goldstein JC et al. P53 induces apoptosis by caspase activation through mitochondrial cytochrome C release. J Biol Chem, 2000, 275(10): 7337-7342.
19. Sitailo LA, Tibudan SS, Denning MF. Activation of caspase-9 is required for UV-induced apoptosis of human keratinocytes. J Biol Chem, 2002, 277(22): 19346-19352.
20. Ettorre A, Andreassi M, Anselmi C etal. Involvement of oxidative stress in apoptosis induced by a mixture of isothiazolinones in normal human keratinocytes. Joural of Investigative Dermatology, 2003, 121 (2): 328-336.
21. Wehrli P, Viard I, Bullani R. etal. Death receptors in cutaneous biology and disease. J Invest Dermatol, 2000, 115(2): 141-148.
22. Takahashi H, Ishida-Yamamoto A, Lizuka H. Ultraviolet B irradiation induces apoptosis of keratinocytes by direct activation of Fas antigen. J Investig Dermatol Symp Proc, 2001, 6(1): 64-68.
23．张晓用，宋天保，caspase-3与细胞凋亡的研究．(Medical Recapitulate)，2002，8(11)：621-623．
24. Saikumar P, Dong Z, Mikhailov V, et al. Apoptosis: definition mechanism, and relevance to disease. Am J Med, 1999, 107(5): 489-506.
25. Hishikawa K, Nakaki T, Fujii T. Connective tissue growth factor induces apoptosis via??caspase-3 in cultured human aortic smooth muscle cells [J]. Eur J Pharmacol, 2000, 392 (1-2): 19-22.
26. Buckley CD, Pilling D, Henriquez NV, et al. RGD peptides induces apoptosis by direct caspase-3 activation [J]. Nature, 1999, 397(6719): 534-539.
27．江继发，魏海明，吴忠豪．检测肿瘤转移指标的临床意义[J]．国外医学：肿瘤学分册，1998，25(2)：82-84．
28．张伟，周韧，陈培辉等．caspase3、bcl-2蛋白在非霍奇金淋巴瘤组织的表达及其与细胞凋亡和增殖的关系．中华病理学杂志，2001，30(6)：434-438．
29．银铎，刘彤，陆景明．caspase-3表达及其与卵巢上皮性肿瘤组织细胞凋亡和增殖的关系．肿瘤防治杂志，2004，11(6)：622-624．
30．姚峰，陈倩，陶琳等．caspase-3基因在肺癌中的表达及其与bcl-2和bax蛋白表达的关系．肿留防治杂志，2004，11(1)：4-6．
31．林静，朱明华．caspase-3与肝细胞癌的细胞凋亡．国外医学·生理、病理科学与临床分册，2002，22(4)：354-356．
32．杨国胜，丁德茂，费正磊．caspase-3与男性泌尿生殖系统疾病．国外医学泌尿系统分册，2003．23(1)：47-49．
33．李南，陈幸生，陈大良．caspase-3在血管瘤组织中的表达及其意义．中国普外基础与临床杂志，2004，11(4)：311-313．
34、樊再雯，万毅新，安真光．bcl-2、ki-67在肺癌组织中的表达及意义．中华结核和呼吸杂志，2002，25(6)：355-356．
35．李红玲，孙秉中，马福成等．非霍奇金淋巴瘤survivin、p53、ki-67的表达．中华血液学杂志．2003，24(10)：543-544．
36．张谊之，张敏，王琳等．尖锐湿疣fas、bcl-2、ki-67的表达与细胞凋亡的关系．华西医大学报，2001，32(1)：73-76．
37．徐咏莲，刘少阳，红大琼．宫颈癌中survivin、ki-67、cyclinB1的表达及临床意义．现代妇产科进展 2004，13(3)：184-186．
38．谢丽，安瑞芳．ki-67在妇科肿瘤中的研究与应用．国外医学妇幼保健分册，2004，15(6)：391-393．
39．王桂芝，卢书信，张克政，caspase-3、bcl-2基因及ki-67在胃癌前病变中的表达及意??义．Chinese Journal of Clinical Health care, 2004, 7(3)：198-200．
40．陈爱兰，邢红梅．增殖性抗原ki-67在慢性外阴营养不良中的表达及中药治疗的影响．山东中医杂志，2003，22(2)：84-85．
41．王艳芳，姚竟，武杰等．91例外阴营养不良患者中药治疗前后性激素变化．中西医结合杂志．1990，10(8)：479-481．
42．余春艳，慧延平，王长军等．外阴白斑与外阴癌性激素受体的测定．中国皮肤性病学杂志，1996，10(5)：271-272．
43．杜惠兰，外阴营养不良患者病变组织中性激素受体的检测．临床与实验病理学杂志，1997，13(2)：145-147．
44. Friedrich EG Jr , Kalra PS. Serum levels of sex hormones in vulvar lichen sclerosus, and the effect of topical testosterone. N Engl J Med, 1984, 310(8): 488-491.
45. Takahashi T, Ogo M, Hibino T. Partial purification and characterization of two distinct types of caspase from human epidermis. J Invest Dermatol, 1998, 111 (3): 367-372.
46. Eckhart L, Ban J, Fischer H, et al. Caspase-14: analysis of gene structure and mRNA expression during keratinocyte differentiation. Biochem Biophys Res Commun, 2000, 277(3): 655-659.
47．袁长青，丁振华．caspase的活化及在细胞凋亡之中的作用[J]．生理科学进展，2002，33(3)：220-224．
48. Droin N, Dubrez L, Eymun B. et al. Upregulation of CASP genes in human tumor cells undergoing etoposide-induced apoptosis. OnCogene. 1998, 16(22): 2885-2894.
49. Krajewska M, Wang HG, Krajewaki S, et al. Immunohistochemical analysis of in vivo patterns of expression of CPP32(Caspase-3), a cell death protease [J]. Cancer Res, 1997, 57(8): 1605-1613.
50. Endl E, Gerdes J . The Ki-67 protein: fascinating forms and an unknown function [J]. Exp Cell Res , 2000, 257(2): 231-237.
51. Scholzen T, Gerdes J . The Ki-67 protein: from the known and the unknown J Cell Physiol, 2000, 182(3): 311-22.
52．李孟达等主编．外阴及阴道肿瘤的诊治．沈阳：辽宁科学技术出版社，1997．35
53. Kagie MJ, Kenter GG, Zomerdijk-Nooijen Y, et al. Human papillomavirus infection in??squamous cell carcinoma of the vulva, in various synchronous epithelial changes and in normal vulvar skin. Gynecol Oncol, 1997, 67(2): 178-183.
54. Akashi K, Nagasaka T,Nakashim N,etal. Squamous cell carctnoma of the vulva and adjacent lesions treated at Nagoya University Hospital from 1965 to 1997. Nagoya J Med Sci,2001,64(3-4):109-121.
55. Vilmer C, Cavelier-Balloy B, Nogues C, etal. Analysis of alterations adjacent to invasive vulvar carcinoma and their relationship with the associated carcinoma: a study of 67cases. Eur J Gynecol Oncol, 1998, 19(1):25-31.
56. Nola M, Blazanovic A, Dotlic S, etal. Invasive squamous cell carcinoma of vulva:prognostic significance of clinicopathologic parameters. Croat Med J. 2005,46(3):436-42.
57. Kagie MJ, Kenter GG, Tollenaar RA, etal. P53 protein overexpression is common and independent of human papillomavirus infection in squamous cell carcinoma of the vulva.Cancer, 1997, 80(7): 1228-1233.
58. Conley LJ, Ellerbrock TV, Bush TJ, etal. HIV-1 infection and risk of vulvovaginal and perianal condylomata acuminata and intraepithelial neoplasia: a prospective cohort study.Lancet. 2002, 359(9301): 108-113.
59. Giaquinto C, Del Mistro A, De Rossi A, etal. Vulvar carcinoma in a 12-year-old girl with vertically acquired human immunodeficiency virus infection. Pediatrics, 2000,106(4):E57.
60. Mosny DS, Brito F, Bender HG. Immunohistochemical investigations of estrogen receptors in normal and neoplastic squamous epithelium of the vulva.J Reprod Med.1990.35(11):1005-1007.
61. Zippel HH, Sander W, Wurz H. Steroid hormone receptors in normal, dystrophic,dysplastic and carcinomatous vulva tissues. Geburtshilfe Frauenheilkd.1985 .45(4):220-225.
62. Jones RW,Rowan DM.Vulvar intraepithelial neoplasia III:a clinical study of the outcome in 113 casls with relation to the later development of invasive vulvar carcinoma.Obstet Gynecol. 1994,84(5):741-745.
63. Ngan HY, Cheung AN, Liu SS, etal. Abnormal expression or mutation of P53 andHPV in vulvar cancer. Eur J Cancer, 1999, 35(3): 481-484.
64. Lopez-Ocejo O, Viloria-Petit A, Bequet-Romero M, et al. Oncogenes and tumor angiogenesis: the HPV-16 E6 on coprotein activates the vascular endothelial growth factor(VEGF) gene promoter in a P53 independent manner. Oncogene, 2000, 19(40): 4611-4620.
65. Engelman DE, Andrade LA, Vassallo J. Human papillomavirus infection and P53 protein expression in vulvar intraepithelial neoplasia and invasive squamous cell carcinoma. Braz J Med Biol Res, 2003, 36(9): 1159-1165.
66. Adamek K, Szczudrawa A, Basta A. Coexistence of VIN and vulvar invasive cancer with intraepithelial neoplasia and invasive carcinoma of the cervix and/or vagina, and HPV infection of the low female genital tract. Ginekol Pol, 2003, 74(9): 657-661.
67. Allen DG, Hutchins AM, Hammet F, et al. Genetic aberrations detected by comparative genomic hybridisation in vulvar cancers. Br J Cancer, 2002, 86(6): 924-928.
68. Madeleine MM, Daling JR, Carter JJ, etal. Cofactors with human papillomavirus in a population-based study of vulvar cancer. J Natl Cancer Inst, 1997, 89(20): 1516-1523.
69. Rolft KJ, Eva KJ, Maclean AB etal. Cell cycle proteins as molecular markers of malignant change in vular lichen sclerosus. Int J Gynecol Cancer, 2001, 11 (2): 113-118
70．陶人川等．增殖细胞核抗原及Ki-67在口腔白斑和鳞癌中的表达及其相关性．实用口腔医学杂志，2002，18(3)：207-210．
71. Winter RN, Kramer A, Borkowski A, et al. Loss of Caspase-1 and caspase-3 protein expression in human prostate cancer. Cancer Res, 2001, 61 (3): 1277-1232.
72. Cheung TH, Chung TK, LoKW, et al. Apotosis-related proteins in cervical intraepithelial neoplasia and squamous cell carcinoma of the cervix. GynecolOncol, 2002, 86(1): 8-14.
73．周菊梅，吴宜林，刘凤英等．FHIT和Ki-67在CIN、宫颈癌中的表达及意义[J]．实用妇产科杂志，2005，21(9)：557-559．
74. Garzetti GG, Ciavattini A, Goteri G, et al. Ki67 antigen immunostaining (MIB Ⅰ monoclonal antibody) in serous ovarian tumors: index of proliferative activity with prognostic significance [J]. Gynecol Oncol, 1995, 56(2): 169-174.75．蔡欣然、黄长玉等．Ki67抗原与TGFβ_1在肝胆管癌中表达及意义．肿瘤防治研究，2000，27(4)：151-152
76. Kyzer S, Gordon PH. Determinative of proliferation activity in colorectal carcinoma using monoclonal untibody Ki67[J]. Dis Colon Rectum, 1997, 40(3): 322-325
77. Matsumoto M, Koniyama K, Okaue M, et al. Predicting tumor metastasis in patients with oral cancer by means of the proliferation marker ki67. J Oral Sci, 1999, 41(2): 53-56.
78．S 司晓辉，金岩等．口腔鳞状细胞癌DNA含量的测定及意义．实用口腔医学杂志，2001．17(5)：403
79．王岩等．上皮内瘤样病变及宫颈癌组织中Ki67、P16的异常表达．Ningxia Med J，2002 Nov，24(11)：99-101
80．靳玉兰等．食道癌前病变及原位癌组织中Ki67、P53、iNos的异常表达．Chin J Oneol，2001 Mar，23(2)：141-1431、Shirer JA, Jr Ray MC. Familial occurrence of lichen sclerosuse atrophicus Case. reports of a mother and daughter . Arch Dermatol, 1987, 123(4): 485-488.
2、王家璧，杨汉英，何祖根等．女阴白色角化病及粘膜白斑与外阴癌的关系[J] 中华皮肤科杂志，1991，(5)：329-331．
3、米纳瓦尔，阿达莱提，拉莱·苏祖克等．口腔鳞癌中HPV16、18型感染和P53蛋白表达的检测研究[J]．中华口腔医学杂志，2001，36(6)：451-453．
4、Bosch FX, Manos MM, Munoz N, et al. Prevalance of human papillomavirus in cervical cancer: a worldwide perspective. J Natl Cancer Inst, 1995, 87(11): 796-802.
5、拉莱·苏祖克，Amy E Noffsinger，买买提艾力等．维吾尔妇女子宫颈癌人类乳头瘤病毒DNA的检测[J]．中华妇产科杂志，1997，32(7)：405-408．
6、Meyers C, Bromberg-White JL, Zhang J, et al. Infectious virions produced from a human papillomavirus type 18/16 genomic DNA chimera. J Virol, 2002, 76(10): 4723-4733.
7、张莉，金晓明，迟月明等．人乳头状瘤病毒感染与外阴营养不良相关性的研究．电子显微学报，2003，22(4)：267-270．
8、Sagerman PM Choi YJ, Hu Y, etal. Human papillomavirus, vulvar dystrophy, and vulvar carcinoma: differential expression of human papillomavirus and vulvar dystrophy in the presence and absence of squamous cell carcinoma of the vulva. Gynecol Oncol, 1996, 61(3): 328-332.
9、Ozkan S, Atabey N, Fetil E, et al. Evidence for Borrelia burgdorferi in morphea and lichen sclerosus. Int J Dermatol, 2000, 39(4): 278-283.
10、王艳芳，姚竟，武杰等．91例外阴营养不良患者中药治疗前后性激素变化．中西医结合杂志，1990，10(8)：479-481．
11、余春艳，慧延平，王长军等．外阴白斑与外阴癌性激素受体的测定．中国皮肤性病学杂志，1996，10(5)：271-27212、郑艾，彭芝兰，曹泽毅．外阴营养不良和恶性肿瘤与雌、孕激素受体关系的研究．华西医大报，2000，31(2)：248-249．
13、杜惠兰，刘敏如，姚先莹等．外阴营养不良患者病变组织中性激素受体的检测，临床与实验病理学杂志，1997，13(2)：145-147．
14、Friedrich EG Jr , Kalra PS. Seru levels of sex hormones in vulvar lichen sclerosus and the effect of topical testosterone. NEryl J Med, 1984, 310(8): 488-491.
15、Coarlson JA, Grabowski R, Chichester P, et al. Comparartive immunophenotypic study of lichen sclerosus: epidermotropricCD57~+ lymphocytes are numerous-implications for pathogenesis. Am J Dermatopathol, 2000, 22(1): 7-16.
16、Azwrdia RM, Luzzi GA, Byren I, et al. Lichen sclerosus in adult men: a study of HLA associations and susceptibility to autoimmune disease. Br-J-Dermatol, 1999, 140(1): 79-83.
17、勾美图，瑞华，付君等．胎盘多肽治疗外阴营养不良94例疗效分析．中国实用妇科与产科杂志，1996，12(6)：369-370．
18、Harringon CI, Dunsmore IR. An investigation into the incidence of auto-immune disorders in patients with lichen sclerosus and atrophicus. Br J Dermatol, 1981, 104(5): 563-566.
19、Scrimin F, Rustja S, Radillo O, et al. Vulvar lichen sclerosus: an immunologic study. Obstet Gynecol, 2000, 95 (1): 147-150.
20、Becagli L, Cadore L. Sjogren' s syndrome and vulvar cancer. Clin Exp Obstet Gynecol. 1987, 14(1) : 69-71.
21、唱捷，何文翠，张建国等．人表皮生长因子与外阴白色病变的关系．中华妇产科杂志，1995．30(4)：209-211．
22、乔丽雅，李亚里，关铮．表皮生长因子受体在外阴营养不良和外阴鳞癌中的表达及其临床意义．解放军医学杂志，2000，25(2)：138-139．
23、杨树君，陈铭，陈诵芬等．中国实验诊断学．2000，4(3)：111-112．
24、Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, bax, that accelerate sprogrammed cell death . Cell, 1993, 74(4): 609-619.
25、欧阳玲，马晓欣，张淑兰．Bcl-2、Bax蛋白表达与外阴营养不良细胞凋亡的相关性研究[J]．中国医科大学学报，2000，29(6)：436-437．26、Scheistroen M, Trope C, Pettersen EO, et al. P53 protein expression in squamous cell carcinoma of the vulva. Cancer, 1999, 85(5): 1133-1138.
27、张式暖，王亚利，王艳华等．食管癌及癌前病变组织中癌基因的表达．中国现代医学杂志，2000，10 (9)：8-9．
28、李远晃，徐志林．胃癌组织中CyclinE和P53蛋白表达的研究．中国现代医学杂志，2002，12 (1)：23-25．
30、尹菊．谷光甘肽、SOD软膏红外线照射治疗外阴营养不良的疗效分析．中国医科大学学报，1998，27(6)：643-644．
31、Berger J, Telser A, Widschwendter M, et al. Expression of retinoic acid receptors in non-neoplastic epithelial disorders of the vulva and normal vulvar skin. Int J Gynecol Pathol, 2000, 19(2): 95-102.
2. Meyers C, Bromberg-White JL, Zhang J , et al. Infectious virions produced from a human papillomavirus type 18/16 genomic DNA chimera J. Virol, 2002, 76(10): 4723-4733.
3．张莉，金晓明，迟月明等．人乳头状瘤病毒感染与外阴营养不良相关性的研究．电子显微学报，2003，22(4)：267-270．
4．尹菊，刘立峰，周仲华．谷光甘肽、SOD软膏红外线照射治疗外阴营养不良的疗效分析．中国医科大学学报，1998，27(6)：643-644．
5．唱捷，何文翠，张建国等．人表皮生长因子与外阴白色病变的关系．中华妇产科杂志，1995，30(4)：209-211．
6．乔丽雅，李亚里，关铮．表皮生长因子受体在外阴营养不良和外阴鳞癌中的表达及其临床意义．解放军医学杂志，2000，25(2)：138-139．
7．杨树君，陈铭，陈诵芬等．外阴营养不良中fas／apol-1表达的生物学意义．中国实验诊断学．2000，4(3)：111-112．
8. Bccagli L, Cadore L. Sjogren's syndrome and vulvar cancer. Clin Exp obstet Gynecol, 1987, 14(1): 69-71.
9. Coarlson JA, Grabowski R, Chichester P, et al. Comparartive immunophenotypic study of lichen sclerosus: epidermotropicCD57~+ lymphocytes are numerous-implications for pathogenesis, Am J Dermatopathol, 2000, 22(1): 7-16.
10．勾美图，瑞华，付君等．胎盘多肽治疗外阴营养不良94例疗效分析．中国实用妇科与产科杂志，1996，12(6)：369-370．
11. Oltvai ZN, Milliman Cl, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a c onserved homolog, Bax, that accelerates programmed cell death. Cell, 1993, 74(4): 609-619.
12．欧阳玲，张淑兰，王小松等．Bcl-2、bax、P53蛋白表达与老年妇女外阴营养不良细??胞凋亡的相关性研究．中国现代医学杂志，2003，13(12)：48-50．
13. Scheistroen M, Trope C, Pettersen EO, et al. P53 protein expression in squamosu cell carcinoma of the vulva. Cancer, 1999, 85(5): 1133-1138.
14．张式暖，王亚利，王艳华等．食管癌及癌前病变组织中癌基因的表达．中国现代医学杂志，2000，10(9)：8-9．
15．李远晃，徐志林．胃癌组织中CyclinE和P53蛋白表达的研究．中国现代医学杂志，2002，12(1)：23-25．
16．宁玉梅，石一复．PCNA在外阴癌及癌前病变中的表达及其临床意义[J]．实用肿瘤杂志，2000，15(4)：234-236．
17．欧阳玲，马晓欣，张淑兰．Bcl-2、Bax蛋白表达与外阴营养不良细胞凋亡的相关性研究[J]．中国医科大学学报 2000，29(6)：436-437．
18. Schuler M, Bossy-Wetzel E, Goldstein JC et al. P53 induces apoptosis by caspase activation through mitochondrial cytochrome C release. J Biol Chem, 2000, 275(10): 7337-7342.
19. Sitailo LA, Tibudan SS, Denning MF. Activation of caspase-9 is required for UV-induced apoptosis of human keratinocytes. J Biol Chem, 2002, 277(22): 19346-19352.
20. Ettorre A, Andreassi M, Anselmi C etal. Involvement of oxidative stress in apoptosis induced by a mixture of isothiazolinones in normal human keratinocytes. Joural of Investigative Dermatology, 2003, 121 (2): 328-336.
21. Wehrli P, Viard I, Bullani R. etal. Death receptors in cutaneous biology and disease. J Invest Dermatol, 2000, 115(2): 141-148.
22. Takahashi H, Ishida-Yamamoto A, Lizuka H. Ultraviolet B irradiation induces apoptosis of keratinocytes by direct activation of Fas antigen. J Investig Dermatol Symp Proc, 2001, 6(1): 64-68.
23．张晓用，宋天保，caspase-3与细胞凋亡的研究．(Medical Recapitulate)，2002，8(11)：621-623．
24. Saikumar P, Dong Z, Mikhailov V, et al. Apoptosis: definition mechanism, and relevance to disease. Am J Med, 1999, 107(5): 489-506.
25. Hishikawa K, Nakaki T, Fujii T. Connective tissue growth factor induces apoptosis via??caspase-3 in cultured human aortic smooth muscle cells [J]. Eur J Pharmacol, 2000, 392 (1-2): 19-22.
26. Buckley CD, Pilling D, Henriquez NV, et al. RGD peptides induces apoptosis by direct caspase-3 activation [J]. Nature, 1999, 397(6719): 534-539.
27．江继发，魏海明，吴忠豪．检测肿瘤转移指标的临床意义[J]．国外医学：肿瘤学分册，1998，25(2)：82-84．
28．张伟，周韧，陈培辉等．caspase3、bcl-2蛋白在非霍奇金淋巴瘤组织的表达及其与细胞凋亡和增殖的关系．中华病理学杂志，2001，30(6)：434-438．
29．银铎，刘彤，陆景明．caspase-3表达及其与卵巢上皮性肿瘤组织细胞凋亡和增殖的关系．肿瘤防治杂志，2004，11(6)：622-624．
30．姚峰，陈倩，陶琳等．caspase-3基因在肺癌中的表达及其与bcl-2和bax蛋白表达的关系．肿留防治杂志，2004，11(1)：4-6．
31．林静，朱明华．caspase-3与肝细胞癌的细胞凋亡．国外医学·生理、病理科学与临床分册，2002，22(4)：354-356．
32．杨国胜，丁德茂，费正磊．caspase-3与男性泌尿生殖系统疾病．国外医学泌尿系统分册，2003．23(1)：47-49．
33．李南，陈幸生，陈大良．caspase-3在血管瘤组织中的表达及其意义．中国普外基础与临床杂志，2004，11(4)：311-313．
34、樊再雯，万毅新，安真光．bcl-2、ki-67在肺癌组织中的表达及意义．中华结核和呼吸杂志，2002，25(6)：355-356．
35．李红玲，孙秉中，马福成等．非霍奇金淋巴瘤survivin、p53、ki-67的表达．中华血液学杂志．2003，24(10)：543-544．
36．张谊之，张敏，王琳等．尖锐湿疣fas、bcl-2、ki-67的表达与细胞凋亡的关系．华西医大学报，2001，32(1)：73-76．
37．徐咏莲，刘少阳，红大琼．宫颈癌中survivin、ki-67、cyclinB1的表达及临床意义．现代妇产科进展 2004，13(3)：184-186．
38．谢丽，安瑞芳．ki-67在妇科肿瘤中的研究与应用．国外医学妇幼保健分册，2004，15(6)：391-393．
39．王桂芝，卢书信，张克政，caspase-3、bcl-2基因及ki-67在胃癌前病变中的表达及意??义．Chinese Journal of Clinical Health care, 2004, 7(3)：198-200．
40．陈爱兰，邢红梅．增殖性抗原ki-67在慢性外阴营养不良中的表达及中药治疗的影响．山东中医杂志，2003，22(2)：84-85．
41．王艳芳，姚竟，武杰等．91例外阴营养不良患者中药治疗前后性激素变化．中西医结合杂志．1990，10(8)：479-481．
42．余春艳，慧延平，王长军等．外阴白斑与外阴癌性激素受体的测定．中国皮肤性病学杂志，1996，10(5)：271-272．
43．杜惠兰，外阴营养不良患者病变组织中性激素受体的检测．临床与实验病理学杂志，1997，13(2)：145-147．
44. Friedrich EG Jr , Kalra PS. Serum levels of sex hormones in vulvar lichen sclerosus, and the effect of topical testosterone. N Engl J Med, 1984, 310(8): 488-491.
45. Takahashi T, Ogo M, Hibino T. Partial purification and characterization of two distinct types of caspase from human epidermis. J Invest Dermatol, 1998, 111 (3): 367-372.
46. Eckhart L, Ban J, Fischer H, et al. Caspase-14: analysis of gene structure and mRNA expression during keratinocyte differentiation. Biochem Biophys Res Commun, 2000, 277(3): 655-659.
47．袁长青，丁振华．caspase的活化及在细胞凋亡之中的作用[J]．生理科学进展，2002，33(3)：220-224．
48. Droin N, Dubrez L, Eymun B. et al. Upregulation of CASP genes in human tumor cells undergoing etoposide-induced apoptosis. OnCogene. 1998, 16(22): 2885-2894.
49. Krajewska M, Wang HG, Krajewaki S, et al. Immunohistochemical analysis of in vivo patterns of expression of CPP32(Caspase-3), a cell death protease [J]. Cancer Res, 1997, 57(8): 1605-1613.
50. Endl E, Gerdes J . The Ki-67 protein: fascinating forms and an unknown function [J]. Exp Cell Res , 2000, 257(2): 231-237.
51. Scholzen T, Gerdes J . The Ki-67 protein: from the known and the unknown J Cell Physiol, 2000, 182(3): 311-22.
52．李孟达等主编．外阴及阴道肿瘤的诊治．沈阳：辽宁科学技术出版社，1997．35
53. Kagie MJ, Kenter GG, Zomerdijk-Nooijen Y, et al. Human papillomavirus infection in??squamous cell carcinoma of the vulva, in various synchronous epithelial changes and in normal vulvar skin. Gynecol Oncol, 1997, 67(2): 178-183.
54. Akashi K, Nagasaka T,Nakashim N,etal. Squamous cell carctnoma of the vulva and adjacent lesions treated at Nagoya University Hospital from 1965 to 1997. Nagoya J Med Sci,2001,64(3-4):109-121.
55. Vilmer C, Cavelier-Balloy B, Nogues C, etal. Analysis of alterations adjacent to invasive vulvar carcinoma and their relationship with the associated carcinoma: a study of 67cases. Eur J Gynecol Oncol, 1998, 19(1):25-31.
56. Nola M, Blazanovic A, Dotlic S, etal. Invasive squamous cell carcinoma of vulva:prognostic significance of clinicopathologic parameters. Croat Med J. 2005,46(3):436-42.
57. Kagie MJ, Kenter GG, Tollenaar RA, etal. P53 protein overexpression is common and independent of human papillomavirus infection in squamous cell carcinoma of the vulva.Cancer, 1997, 80(7): 1228-1233.
58. Conley LJ, Ellerbrock TV, Bush TJ, etal. HIV-1 infection and risk of vulvovaginal and perianal condylomata acuminata and intraepithelial neoplasia: a prospective cohort study.Lancet. 2002, 359(9301): 108-113.
59. Giaquinto C, Del Mistro A, De Rossi A, etal. Vulvar carcinoma in a 12-year-old girl with vertically acquired human immunodeficiency virus infection. Pediatrics, 2000,106(4):E57.
60. Mosny DS, Brito F, Bender HG. Immunohistochemical investigations of estrogen receptors in normal and neoplastic squamous epithelium of the vulva.J Reprod Med.1990.35(11):1005-1007.
61. Zippel HH, Sander W, Wurz H. Steroid hormone receptors in normal, dystrophic,dysplastic and carcinomatous vulva tissues. Geburtshilfe Frauenheilkd.1985 .45(4):220-225.
62. Jones RW,Rowan DM.Vulvar intraepithelial neoplasia III:a clinical study of the outcome in 113 casls with relation to the later development of invasive vulvar carcinoma.Obstet Gynecol. 1994,84(5):741-745.
63. Ngan HY, Cheung AN, Liu SS, etal. Abnormal expression or mutation of P53 andHPV in vulvar cancer. Eur J Cancer, 1999, 35(3): 481-484.
64. Lopez-Ocejo O, Viloria-Petit A, Bequet-Romero M, et al. Oncogenes and tumor angiogenesis: the HPV-16 E6 on coprotein activates the vascular endothelial growth factor(VEGF) gene promoter in a P53 independent manner. Oncogene, 2000, 19(40): 4611-4620.
65. Engelman DE, Andrade LA, Vassallo J. Human papillomavirus infection and P53 protein expression in vulvar intraepithelial neoplasia and invasive squamous cell carcinoma. Braz J Med Biol Res, 2003, 36(9): 1159-1165.
66. Adamek K, Szczudrawa A, Basta A. Coexistence of VIN and vulvar invasive cancer with intraepithelial neoplasia and invasive carcinoma of the cervix and/or vagina, and HPV infection of the low female genital tract. Ginekol Pol, 2003, 74(9): 657-661.
67. Allen DG, Hutchins AM, Hammet F, et al. Genetic aberrations detected by comparative genomic hybridisation in vulvar cancers. Br J Cancer, 2002, 86(6): 924-928.
68. Madeleine MM, Daling JR, Carter JJ, etal. Cofactors with human papillomavirus in a population-based study of vulvar cancer. J Natl Cancer Inst, 1997, 89(20): 1516-1523.
69. Rolft KJ, Eva KJ, Maclean AB etal. Cell cycle proteins as molecular markers of malignant change in vular lichen sclerosus. Int J Gynecol Cancer, 2001, 11 (2): 113-118
70．陶人川等．增殖细胞核抗原及Ki-67在口腔白斑和鳞癌中的表达及其相关性．实用口腔医学杂志，2002，18(3)：207-210．
71. Winter RN, Kramer A, Borkowski A, et al. Loss of Caspase-1 and caspase-3 protein expression in human prostate cancer. Cancer Res, 2001, 61 (3): 1277-1232.
72. Cheung TH, Chung TK, LoKW, et al. Apotosis-related proteins in cervical intraepithelial neoplasia and squamous cell carcinoma of the cervix. GynecolOncol, 2002, 86(1): 8-14.
73．周菊梅，吴宜林，刘凤英等．FHIT和Ki-67在CIN、宫颈癌中的表达及意义[J]．实用妇产科杂志，2005，21(9)：557-559．
74. Garzetti GG, Ciavattini A, Goteri G, et al. Ki67 antigen immunostaining (MIB Ⅰ monoclonal antibody) in serous ovarian tumors: index of proliferative activity with prognostic significance [J]. Gynecol Oncol, 1995, 56(2): 169-174.75．蔡欣然、黄长玉等．Ki67抗原与TGFβ_1在肝胆管癌中表达及意义．肿瘤防治研究，2000，27(4)：151-152
76. Kyzer S, Gordon PH. Determinative of proliferation activity in colorectal carcinoma using monoclonal untibody Ki67[J]. Dis Colon Rectum, 1997, 40(3): 322-325
77. Matsumoto M, Koniyama K, Okaue M, et al. Predicting tumor metastasis in patients with oral cancer by means of the proliferation marker ki67. J Oral Sci, 1999, 41(2): 53-56.
78．S 司晓辉，金岩等．口腔鳞状细胞癌DNA含量的测定及意义．实用口腔医学杂志，2001．17(5)：403
79．王岩等．上皮内瘤样病变及宫颈癌组织中Ki67、P16的异常表达．Ningxia Med J，2002 Nov，24(11)：99-101
80．靳玉兰等．食道癌前病变及原位癌组织中Ki67、P53、iNos的异常表达．Chin J Oneol，2001 Mar，23(2)：141-1431、Shirer JA, Jr Ray MC. Familial occurrence of lichen sclerosuse atrophicus Case. reports of a mother and daughter . Arch Dermatol, 1987, 123(4): 485-488.
2、王家璧，杨汉英，何祖根等．女阴白色角化病及粘膜白斑与外阴癌的关系[J] 中华皮肤科杂志，1991，(5)：329-331．
3、米纳瓦尔，阿达莱提，拉莱·苏祖克等．口腔鳞癌中HPV16、18型感染和P53蛋白表达的检测研究[J]．中华口腔医学杂志，2001，36(6)：451-453．
4、Bosch FX, Manos MM, Munoz N, et al. Prevalance of human papillomavirus in cervical cancer: a worldwide perspective. J Natl Cancer Inst, 1995, 87(11): 796-802.
5、拉莱·苏祖克，Amy E Noffsinger，买买提艾力等．维吾尔妇女子宫颈癌人类乳头瘤病毒DNA的检测[J]．中华妇产科杂志，1997，32(7)：405-408．
6、Meyers C, Bromberg-White JL, Zhang J, et al. Infectious virions produced from a human papillomavirus type 18/16 genomic DNA chimera. J Virol, 2002, 76(10): 4723-4733.
7、张莉，金晓明，迟月明等．人乳头状瘤病毒感染与外阴营养不良相关性的研究．电子显微学报，2003，22(4)：267-270．
8、Sagerman PM Choi YJ, Hu Y, etal. Human papillomavirus, vulvar dystrophy, and vulvar carcinoma: differential expression of human papillomavirus and vulvar dystrophy in the presence and absence of squamous cell carcinoma of the vulva. Gynecol Oncol, 1996, 61(3): 328-332.
9、Ozkan S, Atabey N, Fetil E, et al. Evidence for Borrelia burgdorferi in morphea and lichen sclerosus. Int J Dermatol, 2000, 39(4): 278-283.
10、王艳芳，姚竟，武杰等．91例外阴营养不良患者中药治疗前后性激素变化．中西医结合杂志，1990，10(8)：479-481．
11、余春艳，慧延平，王长军等．外阴白斑与外阴癌性激素受体的测定．中国皮肤性病学杂志，1996，10(5)：271-27212、郑艾，彭芝兰，曹泽毅．外阴营养不良和恶性肿瘤与雌、孕激素受体关系的研究．华西医大报，2000，31(2)：248-249．
13、杜惠兰，刘敏如，姚先莹等．外阴营养不良患者病变组织中性激素受体的检测，临床与实验病理学杂志，1997，13(2)：145-147．
14、Friedrich EG Jr , Kalra PS. Seru levels of sex hormones in vulvar lichen sclerosus and the effect of topical testosterone. NEryl J Med, 1984, 310(8): 488-491.
15、Coarlson JA, Grabowski R, Chichester P, et al. Comparartive immunophenotypic study of lichen sclerosus: epidermotropricCD57~+ lymphocytes are numerous-implications for pathogenesis. Am J Dermatopathol, 2000, 22(1): 7-16.
16、Azwrdia RM, Luzzi GA, Byren I, et al. Lichen sclerosus in adult men: a study of HLA associations and susceptibility to autoimmune disease. Br-J-Dermatol, 1999, 140(1): 79-83.
17、勾美图，瑞华，付君等．胎盘多肽治疗外阴营养不良94例疗效分析．中国实用妇科与产科杂志，1996，12(6)：369-370．
18、Harringon CI, Dunsmore IR. An investigation into the incidence of auto-immune disorders in patients with lichen sclerosus and atrophicus. Br J Dermatol, 1981, 104(5): 563-566.
19、Scrimin F, Rustja S, Radillo O, et al. Vulvar lichen sclerosus: an immunologic study. Obstet Gynecol, 2000, 95 (1): 147-150.
20、Becagli L, Cadore L. Sjogren' s syndrome and vulvar cancer. Clin Exp Obstet Gynecol. 1987, 14(1) : 69-71.
21、唱捷，何文翠，张建国等．人表皮生长因子与外阴白色病变的关系．中华妇产科杂志，1995．30(4)：209-211．
22、乔丽雅，李亚里，关铮．表皮生长因子受体在外阴营养不良和外阴鳞癌中的表达及其临床意义．解放军医学杂志，2000，25(2)：138-139．
23、杨树君，陈铭，陈诵芬等．中国实验诊断学．2000，4(3)：111-112．
24、Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, bax, that accelerate sprogrammed cell death . Cell, 1993, 74(4): 609-619.
25、欧阳玲，马晓欣，张淑兰．Bcl-2、Bax蛋白表达与外阴营养不良细胞凋亡的相关性研究[J]．中国医科大学学报，2000，29(6)：436-437．26、Scheistroen M, Trope C, Pettersen EO, et al. P53 protein expression in squamous cell carcinoma of the vulva. Cancer, 1999, 85(5): 1133-1138.
27、张式暖，王亚利，王艳华等．食管癌及癌前病变组织中癌基因的表达．中国现代医学杂志，2000，10 (9)：8-9．
28、李远晃，徐志林．胃癌组织中CyclinE和P53蛋白表达的研究．中国现代医学杂志，2002，12 (1)：23-25．
30、尹菊．谷光甘肽、SOD软膏红外线照射治疗外阴营养不良的疗效分析．中国医科大学学报，1998，27(6)：643-644．
31、Berger J, Telser A, Widschwendter M, et al. Expression of retinoic acid receptors in non-neoplastic epithelial disorders of the vulva and normal vulvar skin. Int J Gynecol Pathol, 2000, 19(2): 95-102.