2014 8 28 8 Aug 2014 Vol.28 No.8 CHIN J DERM VENEREOL 771 IL-36Ra IL-36Ra DITRA PCR IL36RN IL36RN c. 115 + 6T > C 178 8 IL36RN c. 115 + 6T > C IL36RN c. 115 + 6T > C DITRA DITRA IL36RN R 758. 63 DOI 10. 13735 /j. cjdv. 1001-7089. 2014. 0771 A 1001-7089 2014 08-0771 - 05 Mutation Analysis of IL36RN in A Chinese DITRA Family with Cutaneous Squamous Cell Carcinoma SHU Dan JIN Hong-zhong ZUO Ya-gang YAN Yan Department of Dermatology PUMC Hospital CAMS and PUMC Beijing 100730 China Corresponding author JIN Hong-zhong E-mail jinhongzhong@ 263. net Abstract Objective To report a DITRA family in which the proband developed cutaneous squamous cell carcinoma secondary to DITRA. Methods The blood samples of a Chinese DITRA family and 178 unrelated healthy controls were collected. All the exons and the flanking sequences of IL36RN were amplified by PCR and products analyzed by direct sequencing. Results Histopathological examination of the skin ulcer revealed epithelioid cells with atypia arranged in cords invasive growth of the dermis and mutation analysis of IL36RN revealed homozygous mutation c. 115 + 6T > C in the proband. Meanwhile IL36RN mutation a- nalysis was performed in the unaffected family members and 178 healthy controls. All the unaffected family members and eight healthy controls carried heterozygous c. 115 + 6T > C mutation in IL36RN. Conclusion Homozygous IL36RN c. 115 + 6T > C mutation may be the cause of DITRA and refractory and persistent pustular lesions may be the causes of secondary cutaneous squamous cell carcinoma in the proband. Key words DITRA IL36RN Mutation Cutaneous squamous cell carcinoma generalized pustular psoriasis GPP RA IL36RN DIT- GPP IL36RN IL- 36Ra GPP 1 1. 1 25 7 2011 Marrakchi 1 IL-36Ra 2 DITRA GPP 1 2 1 4-5 4 3 3 ~ 8d 1 20121106110047 201002016 100730 E-mail jinhongzhong@ 263. net
772 8 1d ~ 1e X DITRA 12 A 20mg /d A 100mg /d 21 3 6 A 100mg /d 25mg 3d 1 37kg 3 1f ~ 1g 3 Ⅳ 2 1a 10cm 8cm AE1 /AE3 3 + EMA + CEA - CK7 1b - 2 1c 2 86mm /h C 3kg 1 30. 53mg /LRF HLA-B27 3 PPD + 1. 2 DNA 5 X 5mL DNA 178 ~ Clinical manifestations before treatment X-ray manifestation of feet X-ray manifestation of lower right leg ~ Clinical manifestations after treatment Fig. 1 1 X Clinical and X-ray manifestations of the proband
773 HE staining bar = 1mm HE staining bar = 100μm AE1 /AE3 3 + SP staining bar = 200μm EMA + SP staining bar = 200μm Fig. 3 3 Fig. 2 2 Histopathological and immunohistochemical examinations of the ulcer Pedigree of the family DNA 1. 3 PCR c. 115 + 6T > C 4b ~ IL36RN GenBank NM_012275 5 4d IL36RN c. 115 + 6 T > C 5 2. 24% IL1RN 3 1 PCR 2. 2 IL36RN mrna RT-PCR 2 PCR 271bpmRNA 3 IL36RN 5a ~ 5b IL36RN c. 115 + 6T > C IL-1Ra 1 IL36RN PCR RT-PCR Tab. 1 Primer pairs for IL36RN PCR and RT-PCR amplification DIRA 6 IL1RN 1. 4 PCR RT-PCR IL36RN mrna 1 IL36RN c. 115 + 6T > C RT-PCR TaKaRa RNA RNA cd- NA cdna PCR 1 2 2. 1 DNA 4a UCSC http genome. ucsc. edu / IL36RN 3 IL36RN c. 115 + 6T > C 178 8 Exon /cdna Forward primer Reverse primer Productsize bp 1 5'-AGTGCTTCTGGCGACTTAGG-3' 5'-GGAGAGAGAGGCTGAGTTGG-3' 276 2 5'-CTGACCCCAGACCCAGAATC-3' 5'-AGCTGGACAACGGGTCTATC-3' 261 3 5'-GTTACTTCTGGCACAGTAGG-3' 5'-CACTTTGCTGAGAGGTGTAG-3' 392 4 5'-TCATGACAGCTGCTGAGAAG-3' 5'-AGCTGCCATCAACAGAATCC-3' 386 5. 1 5'-AGATGCTGAGCCTACTGAAG-3' 5'-TCTGACATCAGCACCTCCTC-3' 950 5. 2 5'-TAGAGTCAGGGATCTATGGC-3' 5'-GTGTCCTCTCCTTTTCATAC-3' 940 5. 3 5'-CAAATTCACATCCTTCTTGG-3' 5'-GGGTAAATGAAGGATAGTTG-3' 917 cdna 5'-GTCCTGAGTGGGGCGCTGTG-3' 5'-GAAGTAGAAGTCTGTGATGGG-3' 450
774 RT-PCR 450bpmR- NA 5a 5c 3 GPP IL1RN 7 PsA Rieter GPP DITRA Friedman 8 1 GPP RF HLA-B27 PUVA A 9-10 Gel electrophoresis of amplified IL36RN DNA fragments M DNA Ladder Marker 1 ~ 7 The amplified DNA fragments of IL36RN exon 1-5. 3 Homozygous IL36RN c. 115 + 6T > C mutation Heterozygous IL36RN c. 115 + 6 T > C mutation Wild type of IL36RN Fig. 4 4 PCR Gel electrophoresis and PCR sequencing of amplified IL36RN DNA fragments PsA Rieter IL-1Ra IL1RN Piepkorn 11 1 25 GPP A 4 A A TNF-α IL36RN c. 115 + 6T > C DITRA IL36RN c. 115 + 6T > C IL36RN 3 2 4 IL36RN 155 IL-36Ra 3 10 IL-36Ra /IL- 36R DITRA 4 5 Gel electrophoresis of amplified IL36RN cdna fragments M DNA Ladder Marker 1 Patient 2 Healthy control RT-PCR sequencing in the 8 c. 115 + 6T > C proband PTC premature termination condon RT-PCR sequencing in the healthy control IL-36Ra 5 RT-PCR IL-36 /IL-36R Fig. 5 Gel electrophoresis and PCR sequencing of amplified IL36RN cdna fragments IL36RN c. 115 + 6 T > C 2. 24%
775 psoriasis J. Dermatology 2013 226 1 28-31. 4. 12% 0. 37% IL36RN homozygous deletion of the IL1RN locus J. N Engl J Med c. 115 + 6 T > C DITRA 23 2438-2444. 1 Marrakchi S Guigue P Renshaw BR et al. Interleukin-36-receptor antagonist deficiency and generalized pustular psoriasis J. N Engl J Med 2011 365 7 620-628. 2 Onoufriadis A Simpson MA Pink AE et al. Mutations in IL36RN / IL1F5 are associated with the severe episodic inflammatory skin disease known as generalized pustular psoriasis J. Am J Hum Genet 2011 89 3 432-437. 3 Farooq M Nakai H Fujimoto A et al. Mutation analysis of the IL36RN gene in 14 japanese patients with generalized pustular psoriasis J. hum Mutat 2013 34 1 176-183. 4 Setta-Kaffetzi N Navarini AA Patel VM et al. Rare pathogenic variants in IL36RN underlie a spectrum of psoriasis-associated pustular phenotypes J. J Invest Dermatol 2013 133 5 1366-1369. 5 Abbas O Itani S Ghosn S et al. Acrodermatitis continua of Hallopeau is a clinical phenotype of DITRA evidence that it is a variant of pustular 6 Reddy S Jia S Geoffrey R et al. An autoinflammatory disease due to 2009 360 7 Abbas O Itani S Ghosn S et al. Acrodermatitis continua of Hallopeau is a clinical phenotype of DITRA evidence that it is a variant of pustular psoriasis J. Dermatology 2013 226 1 28-31. 8 Friedman KJ Vollmer RT. Malignant melanoma and squamous cell carcinoma in a patient treated for severe pustular psoriasis J. J Dermatol Surg Oncol 1986 12 12 1290-1294. 9 Pouplard C Brenaut E Horreau C et al. Risk of cancer in psoriasis a systematic review and meta-analysis of epidemiological studies J. J Eur Acad Dermatol Venereol 2013 27 Suppl 3 36-46. 10 Brewer JD Hoverson Schott AR Roenigk RK. Multiple squamous cell carcinomas in the setting of psoriasis treated with etanercept a report of four cases and review of the literature J. Int J Dermatol 2011 50 12 1555-1559. 11 Piepkorn M Kumasaka B Krieger JN et al. Development of human papillomavirus-associated Buschke-L wenstein penile carcinoma during cyclosporine therapy for generalized pustular psoriasis J. J Am Acad Dermatol 1993 29 2 Pt 2 321-325. 2013-10-18 2014-03-11 1 HPV 6 11 13 16 18 30 1 2 2005 1 3 5 4 11 1 42 1 9 1 0. 1cm 0. 1cm HPV 0. 8cm 0. 6cm 1. M. 2010 1 413-418. 2. 1 J. 2005 19 1 58. 3. 5 J. 2 2002 23 1 93. 2 4. 11 J. HPV HPV 2003 13 5 465-470. R 752. 5 + 3 B 1001-7089 2014 08-0775 - 01 DOI 10. 13735 /j. cjdv. 1001-7089. 2014. 0775 1 2 HE 40 650000 2014-02-12 2014-03-17