摘要
【研究目的】(1)研究在眼睑基底细胞癌(Basal Cell Carcinoma,BCC)中NES1基因和蛋白的表达情况，并分析其表达与表型之间的关系，从基因和蛋白水平上研究NES1在BCC的发生发展过程中的作用，以期判断NES1基因是否能作为眼睑肿瘤早期诊断和预后判断的标记物；(2)研究NES1基因外显子3的甲基化情况，以期找到NES1在眼睑BCC中异常表达的可能机制，为眼睑BCC的治疗寻找新的靶点。
【研究方法】本实验共分两大部分：第一部分，我们采用实时荧光定量PCR(Real-time Quantitative PCR)和免疫组化的方法检测了NES1在31例眼睑BCC中的基因表达和蛋白(human kallikrein 10,hk10)的表达情况，并分析其表达与表型之间的关系，我们采用Image-Pro Plus5.0图像分析系统对免疫组化结果进行扫描，定量分析了hk10在不同组眼睑BCC中表达；第二部分，采用甲基化特异性PCR(Methylation Specific
Purpose (1)To investigate the expression of NES1 gene and protein in eyelid basal cell carcinoma (BCC),and analyze the correlation between its expression and the tumorigenic phenotype. In addition, to study the role of NES1 in oncogenesis and progression of eyelid basal cell carcinoma at gene and protein level, and estimate the feasibility as a tumor marker in the early diagnosis and prognosis.(2) In order to find out the mechanisms of loss of NES1 expression, we have studied the methylation condition in exon 3,and expect to find out a new target in the treatment of eyelid BCC.
Methods The experiment has two parts. In the first part, we have studied the expression of NES1 gene and protein in 31 cases of eyelid basal cell carcinoma by using real-time quantitative PCR and immunohistochemistry. and analyzed the correlation between its expression and the tumorigenic phenotype at gene and protein level. We quantificationally analyzed the expression of hk10 in different groups by using Image-Pro Plus5.0 soft. In
引文
1 李凤鸣，主编．眼科全书．人民卫生出版社，1996：1031-1032．
2 孙为荣，牛膺筠，主编．眼科肿瘤学．人民卫生出版社，2004：124-127．
3 Allali J, D'Hermies F, Renard G. Basal cell carcinomas of the eyelids. Ophthalmologica. 2005 Mar-Apr;219(2):57-71.
4 Ducasse A, Pluot M, Gotzamanis A, et al. Rossi P. Factors of recurrence of basal cell carcinomas of the eyelid. J Fr Ophtalmol. 2002 May;25(5):512-516.
5 Luo L, Herbrick JA, Scherer SW, et al. Structural characterization and mapping of the normal epithelial cell-specific1 gene. Biochem Biophys Res Commun, 1998,247 (3):580-586.
6 Roman-Gomez J, Jimenez-Velasco A, Agirre X, et al. The normal epithelial cell-specific 1 (NES1) gene, a candidate tumor suppressorgene on chromosome 19q13.3-4, is downregulated by hypermethylation in acutelymphoblastic leukemia. Leukemia. 2004 Feb; 18(2):362-365.
7 Li B, Goyal J, Dhar S, et al. CpG methylation as a basis for breast tumor-specific loss of NES1/kallikrein 10 expression. Cancer Res.2001 Nov1;61(21):8014-8021.
8 Goyal J, Smith KM, Cowan JM,et al. The role for NES1 serine protease as a novel tumor suppressor. Cancer Res. 1998 Nov 1;58(21):4782-4786.
9 Liu XL, Wazer DE, Watanabe K, et al. Identification of a novel serine protease-like gene, the expression of which is down-regulated during breast cancer progression. Cancer Res. 1996 Jul 15;56(14):3371-3379.
10 Petraki CD, Karavana VN, Luo LY, et al. Human kallikrein 10 expression in normal tissues by immunohistochemistry. J Histochem Cytochem,2002, 50(9):1247-1261
11 Shvartsman HS, Lu KH, Lee J , et al .Overexpression of kallikrein 10 in epithelial ovarian carcinomas. Gynecol Oncol, 2003, 90(1 ):44-50.
12 Lou LY, Katsaros D, Scorilas A, et al. Prognostic value of human kallikrein 10 expression in epithelial ovarian carcinoma. Clin Cancer Res, 2001,7(8):2372-2379.
13 Luo LY, Rajpert-De Meyts ER, Jung K, et al. Expression of the normal epithelial cell-specific 1 (NES1; KLK10) candidate tumour suppressor gene in normal and malignant testicular tissue. Br J Cancer, 2001, 85(2):220-224.
14 Kenneth J. Livak and Thomas D. Schmittgen. Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2-△△CT Method. METHODS ,2001,25:402-408.
15 Higuchi R, Fockler C, etal. Kinetic PCR analysis: real-time monitoring of DNA amplification reactions. Biotechnology, 1993,11(9): 1026-1030.
16 Becker K., D. Pan and C.B.Whitely. Real-time quantitative polymerase chain reaction to assess gene transfer. Hum. Gene Ther, 1999, 10: 2559-2566.
17 Wang X. X. Li, etal. Application of real-time polymerase chain reaction to quantitate induced expression of interleukin-1 beta mRNA in ischemic brain tolerance. Neurosci.Res, 2000, 59(2): 238-46.
18 Actor JK, Limor JR, Hunter RL. A flexible bioluminescent-quantitive polymerase reaction assay for analysis of competitive PCR amplicons. J Clin Lab Anal, 1999,13(1): 40-47.
19 Borgono CA, Michael IP, Diamandis EP, Human tissue kallikreins: physiologic roles and applications in cancer. Mol Cancer Res,2004,2(5): 257-280.
20 Dhar S, Bhargava R, Yunes M, et al. Analysis of normal epithelial cells pecific-l( NESl)/kallikrein 10m RNA expression by in situ hybridization, a novel marker for breast cancer. Clin Cancer Res,2001,7(11):3393-3398.
21 Yunes MJ, Neuschatz AC, Bornstein LE, et a 1. Loss of expression of the putative tumor suppressor NESl gene in biopsy-proven ductal carcinoma in situ predicts for invasive carcinoma at definitive surgery. Int J Radiat Oncol Biol Phys,2003,56(3):653-657.
22 Luo LY, Katsaros D ,Scorilas A ,et al.The serum concentration of human kallikrein 10 represents a novel biomarker for ovarian cancer diagnosis and prognosis. Cancer Res,2003,63(4);807-811.
23 Oikonomopoulou K, Scorilas A, Michael IP, et al. Kallikreins as Markers of Disseminated Tumour Cells in Ovarian Cancer - A Pilot Study. Tumour Biol. 2006 Mar 24;27(2):104-114.
24 Sidiropoulos M, Pampalakis G, Sotiropoulou G, et al. Downregulation of human kallikiein 10 (KLK10/NES1) by CpG island hypermethylation in breast, ovarian and prostate cancers. Tumour Biol. 2005 Nov-Dec;26(6): 324-336.
25 Rawlinson A, Elcock C, Cheung A,et al. An in-vitro and in-vivo methodology study of alveolar bone measurement using extra-oral radiographic alignment apparatus, Image Pro-Plus software and a subtraction programme. J Dent. 2005 Oct;33(9):781-788.
26 Blatt RJ, Clark AN, Courtney J, et al. Automated quantitative analysis of angiogenesis in the rat aorta model using Image-Pro Plus 4.1. Comput Methods Programs Biomed. 2004 Jul;75(1):75-79.
27 Xavier LL, Viola GG, Ferraz AC, et al. A simple and fast densitometric method for the analysis of tyrosine hydroxylase immunoreactivity in the substantia nigra pars compacta and in the ventral tegmental area. Brain Res Brain Res Protoc. 2005 Dec;16(1-3):58-64.
28 Obenauer-Kutner LJ, Halperin R, Ihnat PM, et al. Use of an automated image processing program to quantify recombinant adenovirus particles.Microsc Microanal. 2005 Feb;11(1):37-41.
1 Wu Ct, Morris JR. Genes, genetics, and epigenetics: a correspondence. Science. 2001 Aug 10;293(5532):1103-1105.
2 Imai K, Toyota M, Sato H, et al. DNA methylation and carcinogenesis. Nippon Naika Gakkai Zasshi. 2006 Feb 10;95(2):362-367
3 PazMF, Yaya-Tur R, Rojas-Marcos I, et al. CpG island hypermethylation of the DNA repair enzyme methyltransferase predicts response to temozolomide in primary gliomas. Clin Cancer Res, 2004,10:4933-4938.
4 Roman-Gomez J, Jimenez-Velasco A, Agirre X, et al. The normal epithelial cell-specific 1 (NES1) gene, a candidate tumor suppressorgene on chromosome 19q13.3-4, is downregulated by hypermethylation in acutelymphoblastic leukemia. Leukemia. 2004 Feb;18(2):362-365.
5 Li B, Goyal J, Dhar S, et al. CpG methylation as a basis for breast tumor-specific loss of NES1/kallikrein 10 expression. Cancer Res.2001 Nov1;61(21):8014-8021.
6 Sidiropoulos M, Pampalakis G, Sotiropoulou G, et al. Downregulation of human kallikiein 10 (KLK10/NES1) by CpG island hypermethylation in breast, ovarian and prostate cancers. Tumour Biol. 2005 Nov-Dec;26(6): 324-336
7 Ahmad I, Rao DN. Chemistry and biology of DNA methyltransferases. Crit Rev Biochem MolBiol, 1996,31: 361-380.
8 Vertino PM, Yen RW, Gao J , et al. De novo methylation of CpG island sequences in human fibroblasts overexpressing DNA(cytosine-5) -Meth- yltransferasel Mol Cell Biol ,1996 ,16 (8) :4555
9 G. Strathdee and R. Brown. Aberrant DNA methylation in cancer: potential clinical interventions. Expert Rev Mol Med, 2002,3: 1-17.
10 Herman JG, Baylin SB. Gene silencing in cancer in association with promoter hypermethylation. N Engl J Med,2003,349(21):2042-2054
11 Ehrlich M, Wang RY. 5-methylcytosine in eukaryotic DNA.Science, 1981,212(4501):1350-1357
12 Jones PA, Gonzalgo ML, Tsutsumi M, et al. DNA methylation in bladder cancer Eur Urol. 1998;33 Suppl 4:7-8.
13 周玉球．DNA甲基化分析技术的研究进展．国外医学遗传学分册2001，24(6)：303-308
14 Herman JG, Graft JR, Myohanen S, et al. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9821-9826.
15 武立鹏，朱卫国．DNA甲基化的生物学应用及检测方法进展．中华检验医学杂志，2004，27(7)：468-474．
16 朱燕．DNA甲基化的分析与状态检测．现代预防医学，2005，32(9)：1070-1073．
17 黄庆，府伟灵．DNA甲基化分析技术．中国实验诊断学，2005，9(2)：304-306
18 Uchida T, Ohashi H, Aoki E, et al. Clonality analysis by methylation-specific PCR for the human androgen-receptor gene(HUMARA-MSP). Leukemia. 2000 Jan;14(1):207-212.
19 Goessl C, Muller M, Heicappell R, et al. Methylation-specific PCR for detection of neoplastic DNA in biopsy washings.J Pathol. 2002 Mar; 196(3):331-4.
20 Derks S, Lentjes MH, Hellebrekers DM, et al. Methylation-specific PCR unraveled.Cell Oncol. 2004;26(5-6):291-299.
21 Cottrell SE, Laird PW. Sensitive detection of DNA methylation.Ann N Y Acad Sci. 2003 Mar;983:120-130.
22 Li B, Goyal J, Dhar S, et al. CpG methylation as a basis for breast tumor-specific loss of NES1/kallikrein 10 expression. Cancer Res.2001; 61(21):8014-8021.
23 Yunes MJ, Neuschatz AC, Bornstein LE, et al. Loss of expression of the putative tumor suppressor NESl gene in biopsy-proven ductal carcinoma in situ predicts for invasive carcinoma at definitive surgery. Int J Radiat Oncol Biol Phys. 2003 Jul 1;56(3):653-657.
24 Roman-Gomez J, Jimenez-Velasco A, Agirre X, et al. The normal epithelial cell-specific 1 (NESl) gene, a candidate tumor suppressorgene on chromosome 19q13.3-4, is downregulated by hypermethylation in acutelymphoblastic leukemia. Leukemia. 2004 Feb;18(2):362-365.
25 Sidiropoulos M, Pampalakis G, Sotiropoulou G, et al. Downregulation of human kallikrein 10 (KLK10/NES1) by CpG island hypermethylation in breast, ovarian and prostate cancers. Tumour Biol. 2005 Nov-Dec;26(6): 324-336
26 Luo LY, Yousef G, Diamandis EP. Human tissue kallikreins and testicular cancer. APMIS. 2003 Jan;111(1):225-232; discussion 232-233.
27 Luo LY, Katsaros D ,Scorilas A ,et al.The serum concentration of human kallikrein 10 represents a novel biomarker for ovarian cancer diagnosis and prognosis. Cancer Res,2003,63(4);807-811.
28 Gore SD. Combination therapy with DNA methyltransferase inhibitors in hematologic malignancies.Nat Clin Pract Oncol. 2005 Dec;2 Suppl 1:S30-35.
29 Hess JL. Mechanisms of transformation by MLL. Crit Rev Eukaryot Gene Expr. 2004;14(4):235-254.
1 Petraki CD, Karavana VN, Luo LY, et al.Human kaUikrein 10 expression in normal tissues by immunohistochemistry. J Histochem Cytochem,2002,50(9):1247-1261
2 Roman-Gomez J, Jimenez-Velasco A, Agirre X, et al. The normal epithelial cell-specific 1 (NES1) gene, a candidate tumor suppressorgene on chromosome 19q13.3-4, is downregulated by hypermethylation in acutelymphoblastic leukemia. Leukemia. 2004 Feb;18(2):362-365.
3 Li B, Goyal J, Dhar S, et al. CpG methylation as a basis for breast tumor-specific loss of NES1/kallikrein 10 expression. Cancer Res.2001 Nov1;61(21):8014-8021.
4 Sidiropoulos M, Pampalakis G; Sotiropoulou G, et al. Downregulation of human kallikrein 10 (KLK10/NES1) by CpG island hypermethylation in breast, ovarian and prostate cancers. Tumour Biol. 2005 Nov-Dec;26(6): 324-336
5 Liu XL, Wazer DE, Watanabe K, et al. Identification of a novel serine protease-like gene, the expression of which is down-regulated during breast cancer progression. Cancer Res. 1996 Jul 15;56(14):3371-3379.
6 Borgono CA, Michael IP, Diamandis EP, Human tissue kallikreins: physiologic roles and applications in cancer. Mol Cancer Res,2004,2(5): 257-280.
7 Luo L, Herbrick JA, Scherer SW, et al. Structural characterization and mapping of the normal epithelial cell-specific1 gene. Biochem Biophys Res Commun,1998,247 (3):580-586.
8 Yousef GM, Luo LY, Diamandis EP. Identification of novel human kallikrein-like genes on chromosome 19q13.3-q13.4. Anticancer Res. 1999 Jul-Aug;19(4B):2843-2852.
9 Goyal J, Smith KM, Cowan JM,et al. The role for NES1 serine protease as a novel tumor suppressor. Cancer Res. 1998 Nov 1;58(21):4782-4786.
10 Luo LY, Grass L, Howarth DJ, et al. Immunofluorometric assay of human kallikrein 10 and its identification in biological fluids and tissues. Clin Chem. 2001 Feb;47(2):237-246.
11 Luo LY,Grass L, Diamandis EP. The normal epithelial cell-specific1 ( NES1 )gene is up-regulated by steroid hormones in the breast carcinoma cell line BT-474.Anticancer Res,2000.20( 2A): 981-986.
12 Luo LY, Grass L, Diamandis EP. Steroid hormone regulation of the human kallikrein 10 (KLK10) gene in cancer cell lines and functional characterization of the KLK10 gene promoter. Clin Chim Acta. 2003 Nov;337(1-2):115-26.
13 Yousef GM, Scorilas A, Diamandis EP. Genomic organization, mapping, tissue expression, and hormonal regulation of trypsin-like serine protease (TLSP PRSS20), a new member of the human kallikrein gene family. Genomics. 2000 Jan 1;63(1):88-96.
14 Diamandis EP, Yousef GM, Luo LY, et al. The new human kallikrein gene family: implications in carcinogenesis. Trends Endocrinol Metab. 2000 Mar;11(2):54-60.
15 Dhar S, Bhargava R, Yunes M, et al. Analysis of normal epithelial cells pecific-1( NES1)/kallikrein 10m RNA expression by in situ hybridization, a novel marker for breast cancer. Clin Cancer Res,2001,7( 11):3393-3398.
16 Yunes MJ, Neuschatz AC, Bornstein LE, et a 1. Loss of expression of the putative tumor suppressor NES1 gene in biopsy-proven ductal carcinoma in situ predicts for invasive carcinoma at definitive surgery. Int J Radiat Oncol Biol Phys,2003,56(3):653-657.
17 Luo LY, Diamandis EP, Look MP, et al.Higher expression of human Kallikrein 10 in breast cancer tissue predicts tamoxifen resistance.Br J Cancer, 2002,86(11):1790-1796.
18 Shvartsman HS, Lu KH, Lee J , et al .Overexpression of kallikrein 10 in epithelial ovarian carcinomas. Gynecol Oncol, 2003, 90(1 ):44-50.
19 Luo LY, Katsaros D, Scorilas A,et al. The serum concentration of human kallikrein 10 represents a novel biomarker for ovarian cancer diagnosisand prognosis. Cancer Res. 2003 Feb 15;63(4):807-811.
20 Rosen DG, Wang L, Atkinson JN, et al. Potential markers that complement expression of CA125 in epithelial ovarian cancer. Gynecol Oncol. 2005 Nov;99(2):267-277.
21 Luo LY, Rajpert-De Meyts ER, Jung K, et al. Expression of the normal epithelial cell-specific 1 (NES1; KLK10) candidate tumour suppressor gene in normal and malignant testicular tissue. Br J Cancer, 2001, 85(2):220-224.
22 Luo LY, Yousef G, Diamandis EP. Human tissue kallikreins and testicular cancer.APMIS. 2003 Jan;111(1):225-32
23 Sidiropoulos M, Pampalakis G, Sotiropoulou G, et al. Downregulation of human kallikrein 10 (KLK10/NES1) by CpG island hypermethylation in breast, ovarian and prostate cancers. Tumour Biol. 2005 Nov-Dec;26(6): 324-336.
24 Roman-Gomez J, Jimenez-Velasco A, Agirre X, et al. The normal epithelial cell-specific 1 (NES1) gene, a candidate tumor suppressorgene on chromosome 19q13.3-4, is downregulated by hypermethylation in acutelymphoblastic leukemia. Leukemia. 2004 Feb;18(2):362-365.
25 Dasgupta S, Tripathi PK, Qin H, Identification of molecular targets for immunotherapy of patients with head and neck squamous cell carcinoma. Oral Oncol. 2006 Mar;42(3):306-316.
26 Abmad I, Rao DN. Chemistry and biology of DNA methyltransferases. Crit Rev Biochem MolBiol, 1996, 31: 361-380.
27 Wu Ct, Morris JR. Genes, genetics, and epigenetics: a correspondence. Science. 2001 Aug 10;293(5532):1103-1105.
28 Li B, Goyal J, Dhar S, et al. CpG methylation as a basis for breast tumor-specific loss of NES1/kallikrein 10 expression. Cancer Res.2001 Nov1;61(21):8014-8021.
29 Herman JG,Baylin SB. Gene silencing in cancer in association with promoter hypermethylation. N Engl J Med,2003,349(21):2042-2054
30 武立鹏，朱卫国．DNA甲基化的生物学应用及检测方法进展．中华检验医学杂志，2004，27(7)：468-474．
31 Imai K, Toyota M, Sato H, et al. DNA methylation and carcinogenesis. Nippon Naika Gakkai Zasshi. 2006 Feb 10;95(2):362-367.
32 Kim DH, Shim JS, Kwon HJ. Coordinated transcriptional regulation of calmegin,a testis-specific molecular chaperon, by histone deacetylase and CpG methyltransferase.Exp Mol Med. 2005 Oct 31;37(5):492-496.
33 Enright BP, Sung LY, Chang CC, et al. Methylation and acetylation characteristics of cloned bovine embryos from donor cells treated with 5-aza-2'-deoxycytidine.Biol Reprod. 2005 Apr;72(4):944-948.
34 Gore SD. Combination therapy with DNA methyltransferase inhibitors in hematologic malignancies.Nat Clin Pract Oncol. 2005 Dec;2 Suppl 1:S30-35.
1 Allali J, D'Hermies F, Renard G. Basal cell carcinomas of the eyelids. Ophthalmologica. 2005 Mar-Apr;219(2):57-71.
2 Dazard JE, Gal H,Amariglio N, et al.Genome-wide comparison of human keratinocyte and squamous cell carcinoma response to UVB irradiation: implication for skin and epithelial cancer.Oncogene,2003;22:2993-3006.
3 Saari KM, Paavilainen V, Tuominen J, et al. Epidemiology of basal cell carcinoma of the eyelid in south-western Finland. Graefes Arch Clin Exp Ophthalmol. 2001 Mar;239(3):230-233.
4 刘秉文，陈俊杰主编．医学分子生物学，中国协和医科大学出版社，2000，472-499．
5 Bishop JM. Molecular themes in oncogenesis. Cell,1991,64:235-248.
6 Hunter T. Oncoprotein networks. Cell,1997,88:333-346.
7 Weinberg RA.Tumor suppressor genes.Science,1991,254:1138-1145.
8 孙为荣，牛膺筠，主编．眼科肿瘤学．人民卫生出版社，2004：124-127．
9 李凤鸣，主编．眼科全书．人民卫生出版社，1996：1031-1032．
10 Spencer WH.Ophalmic pathology. Philalphia WH Saunders Company, 1986.2169-2177.
11 周云龙．基底细胞癌生物学和癌发生的研究进展．国外医学皮肤病学分册，1989，15：353
12 Amstad PA, Cerutti PA. Ultraviolet-B-light-induced mutagenesis of C-H-ras codons 11 and 12 in human skin fibroblasts.Int J Cancer. 1995 Sep 27;63(1):136-139.
13 Campbell C, Quinn AG, Rees JL. Codon 12 Harvey-ras mutations are rare events in non-melanoma human skin cancer.Br J Dermatol. 1993 Feb;128(2):111-114.
14 Kikuchi A, Amagai M, Hayakawa K, et al. Association of EGF receptor expression with proliferating cells and of ras p21 expression with differentiating cells in various skin tumours.Br J Dermatol. 1990 Jul;123(1):49-58.
15 Hussein MR. Ultraviolet radiation and skin cancer: molecular mechanisms.J Cutan Pathol. 2005 Mar;32(3):191-205.
16 Barshack I, Goldberg I, Davidson B, et al. Expression of rasGTPase activating protein in basal cell carcinoma of the skin.Mod Pathol. 1998 Mar;11(3):271-275.
17 Nakagawa K, Yamamura K, Maeda S, et al. bcl-2 expression in epidermal keratinocytic diseases.Cancer. 1994 Sep 15;74(6):1720-1724.
18 Verhaegh ME, Sanders CJ, Arends JW, et al. Expression of the apoptosis-suppressmg protein Bcl-2 in non-melanoma skin cancer.Br J Dermatol. 1995 May;132(5):740-744.
19 Smoller BR, Van de Rijn M, Lebrun D,et al. bcl-2 expression reliably distinguishes trichoepitheliomas from basal cell carcinomas.Br J Dermatol. 1994 Jul;131(1):28-31.
20 Bozdogan O, Erkek E, Atasoy P, et al. Bcl-2-related proteins, alpha-smooth muscle actin and amyloid deposits in aggressive and non-aggressive basal cell carcinomas .Acta Derm Venereol. 2002;82(6):423-427.
21 Niu Y, Zhou Z, Liu F, et al. Expression of P16 protein and Bcl-2 protein in malignant eyelid tumors.Chin Med J (Engl). 2002 Jan;115(1):21-25.
22 Takahashi S, Pearse AD, Marks R. Expression of c-fos proto-oncogene mRNA in non-melanoma skin cancer.J Dermatol Sci. 1994 Feb;7(1):54-62.
23 Urabe A, Nakayama J, Taniguchi S, et al. Expression of the c-fos oncogene in chemically-induced mouse tumours and in human skin tumours.J Pathol. 1992 Nov;168(3):281-286.
24 Ro YS. Oncogene interaction in basal cell carcinomas of human skin.J Korean Med Sci. 1995 Apr;10(2):85-92.
25 Bonifas JM, Pennypacker S, Chuang PT, et al. Activation of expression of hedgehog target genes in basal cell carcinomas.J Invest Dermatol. 2001 May;116(5):739-742.
26 Maguire HC Jr, Jaworsky C, Cohen JA. et al. Distribution of neu (c-erbB-2) protein in human skin.J Invest Dermatol. 1989 Jun;92(6):786-790.
27 Liu B, Zhang H, Li S, et al. The expression of c-erbB-1 and c-erbB-2 oncogenes in basal cell carcinoma and squamous cell carcinoma of skin.Chin Med Sci J. 1996 Jun;11(2):106-109.
28 Ahmed NU, Ueda M, Ichihashi M. Increased level of c-erbB-2/neu/ HER-2 protein in cutaneous squamous cell carcinoma.Br J Dermatol. 1997 Jun;136(6):908-912.
29 ontironi R, Mazzucchelli R, Stramazzotti D,et al. Basal cell hyperplasia and basal cell carcinoma of the prostate: a comprehensive review and discussion of a case with c-erbB-2 expressionJ Clin Pathol. 2005 Mar;58(3):290-296.
30 Lebeau S, Masouye I, Berti M, et al. Comparative analysis of the expression of ERBIN and Erb-B2 in normal human skin and cutaneous carcinomas.Br J Dermatol. 2005 Jun;152(6):1248-1255.
31 Kolar Z, Geierova M, Bouchal J,et al. Immunohistochemical analysis of the biological potential of odontogenic keratocystsJ Oral Pathol Med. 2006 Feb;35(2):75-80.
32 Rajabi MA, Rajabi P, Afshar-Moghaddam N. Determination of p53 expression in basal cell carcinoma tissues and adjacent nontumoral epidermis from sun-exposed areas of the head and neck. Arch Iran Med. 2006 Jan;9(1):46-48.
33 Reszec J, Sulkowski S. The expression of P53 protein and infection of human papilloma virus in conjunctival and eyelid neoplasms.Int J Mol Med. 2005 Oct;16(4):559-564.
34 Tsai KY, Tsao H. The genetics of skin cancer.Am J Med Genet C Semin Med Genet. 2004 Nov 15;131C(1):82-92.
35 Ragge NK, Salt A, Collin JR, et al. Gorlin syndrome: the PTCH gene links ocular developmental defects and tumour formation.Br J Ophthalmol. 2005 Aug;89(8):988-991.
36 Hong Zhang, Xiao Li Ping, Patricia K.et al. Role of PTCH and p53 Genes in Early-Onset Basal Cell Carcinoma. American Journal of Pathology. 2001;158:381-385.
37 Gailani.M.R and Bale.A.E. Human patched (PTCH) mRNA is overexpressed consistently in tumor cells of both familial and sporadic basal cell carcinoma. Cancer Res. 1997 Jun 15;57(12):2336-2340.
38 Soufir N, Moles JP, Vilmer C,et al. P16 UV mutations in human skin epithelial tumors.Oncogene. 1999 Sep 23;18(39):5477-5481.
39 Chang TG, Wang J, Chen LW,et al. Loss of expression of the p16 gene is frequent in malignant skin tumors.Biochem Biophys Res Commun. 1997 Jan 3;230(1):85-88.
40 Kanitakis J, Euvrard S, Bourchany D, et al. Expression of the nm23 metastasis-suppressor gene product in skin tumorsJ Cutan Pathol. 1997 Mar;24(3):151-156.
41 Ro YS, Jeong SJ. Expression of the nucleoside diphosphate kinase in human skin cancers: an immunohistochemical study.J Korean Med Sci. 1995 Apr;10(2):97-102.