Determination of HPV by PCR and immunohistochemistry in cervival invasive squamous cell carcinoma

Yıl 2006, Cilt: 37 Sayı: 3, 53 - 60, 01.04.2006

Raci Güney Sevinç Hallaç Keser Nimet Karadayı Sibel Sensu Dilek Yavuzer Aylin Ege Gül Nagehan Özdemir Barışık

Öz

Many epidemiological, clinicopaihological and molecular studies have shown the role of human papilloma virus (HPV) in the pathogenesis of the cervical carcinoma precursor lesions. Polimerase chain reaction (PCR), is very sensitive method for its establishment. Another method to identify HPV in cervival samples is immunohistochemistry (IHC). In this study, we aimed to determine HPV incidence in cervical invasive squamous cell carcinoma (ISCC) by using PCR and IHC and to investigate the correlation between HPV types and histological types. In our pathology department, 26 cases are investigated for HPV presence and types by PCR method. They are evaluated for HPV presence immunohistochemically by BSA-peroxidase (DAB) method. By PCR we found HPV 16 in 23%, 18 in 11.5%, 33 in 4%, 16+33 (mixt infection) in 4% of the cases. We didn't show HPV 6 and 11 in any of the cases. HPV positivity was 30.5%, immunohistochemically. In 5 of the 15 cases that were found to be negative by PCR, HPV was shown by IHC. The percentage of the cases proven to be HPV positive by PCR and/or IHC was 61.5%. When we compared the histological types with HPV types established by PCR, 18% of large cell keratinizing carcinoma (LCKC) was HPV 16 (+), 18% was 18 (+), 9% was 33 (+), 9% was 16+33 (+) and 46% was HPV(-); whereas 13% of large cell non-keratinizing carcinoma (LCNKC) was HPV 16(+), 18% was 18 (+), 69% was HPV(-) and 50% of small cell carcinoma was HPV 16(+), 50% was HPV(-). When we evaluated the histological type distribution oflHC-proven HPV positive cases, 37.5% was LCKC and 62.5% was LCNKC. We find the role and incidence of HPV in cervical carcinoma cases in our country comparable to global evidence; we conclude with the importance of screening programmes.

Anahtar Kelimeler

Cervix Uteri, Uterine Cervical Neoplasms, Human papillomavirus 6, Human papillomavirus 16, Human papillomavirus 18, Human papillomavirus 11, Immunohistochemistry, Carcinoma, Squamous Cell

Serviksin invaziv skuamöz hücreli karsinomlarında HPV'nin PCR ve immünohistokimyasal yöntemlerle belirlenmesi

Öz

Birçok epidemiyolojik klinikopatolojik ve moleküler çalışmalar çoğu servikal kanser prekürsör lejyonlarının patogenezinde human papilloma virüsün (HPV) varlığını ortaya koymuştur. Moleküler biyolojik tekniklerden polimeraz zincir reaksiyonu (PCR) oldukça duyarlı bir yöntemdir. Servikse ait örneklerde HPV yi belirlemede kullanılan bir başka yöntem de immünohistokimyasal (İHK) yöntemdir. Çalışmamızda serviksin invaziv skuamöz hücreli karsinomlarmda (İSHK), PCR ve İHK yöntemi kullanarak HPV insidansını belirlemekve bulunan HPV tipleri ile histolojik tip arasındaki korelasyonu incelemeyi amaçladık. Kliniğimizde İSHK tanısı almış 26 vaka PCR yöntemiyle hem HPV varlığı hem de HPV tipleri açısından; ayrıca İHK olarak HPV varlığı yönünden BSA-peroksidaz (DAB) yöntemiyle incelendi. Olgularımızda PCR yöntemiyle HPV 16 yi %23, HPV18'i %U.5, HPV33'ü % 4, HPV16+33'ü (mikst enfeksiyon) %4 oranında belirledik. HPV 6 ve IVi hiçbir olguda tespit etmedik. 26 olgumuzda İHK yöntemle % 30.5 oranında HPV pozitivitesi saptadık. PCR ile (-) bulduğumuz 15 olgudan 5'inde İHK ile HPV tespit ettik. PCR ve/veya İHK yöntemiyle HPV pozitivitesi saptadığımız olguların oranı %61.5' di. Histolojik tipleri PCR yöntemi ile bulduğumuz HPV tipleri ile karşılaştırdığımızda büyük hücreli keratinize karsinomların (BHKK) % 18'ni HPV 16 (+), % 18'ni HPV 18 (+), % 9 'nu HPV33 (+), %9'nu HPV 16+33 (+), % 46'sını HPV(-) tespit ettik. Büyük hücreli nonkeratinize karsinomların (BHNKK) % 13'üHPV16(+), %18' i HPV 18 (+), % 69'u HPV(-); küçük hücreli karsinomların (KHK) % 50'si HPV16(+), % 50'si HPV(-) di. İHK olarak pozitif olguların histolojik tip dağılımını incelediğimizde % 37.5'ni BHKK %62.5'ni BHNKK olarak belirledik. Yaptığımız çalışmada servikal karsinomlarda HPV'nin rolü ve insidansının dünya çapındaki bulgularla kesin benzerlikler gösterdiğini ve tarama programlarının önemini vurgulamaya çalıştık.

Anahtar Kelimeler

Serviks uteri, Uterin servikal neoplazmlar, İnsan papillomavirüs 6, İnsan papillomavirüs 16, İnsan papillomavirüs 18, İnsan papillomavirüs 11, İmmünohistokimya, Karsinom, yassı hücreli

Kaynakça

• Huang LW, Chao SL, Chen PH, et al. Multiple HPV genotypes in cervical carcinomas: improved DNA detection and typing in archival tissues. Journal of Clinical Virology 2004; 29: 271-276
• Kaufmann AM, Stern PL, Rankin EM, et al
• Safety and immunogenicity of TA-HPV, a recombinant vaccinia virus expressing modified human papillomavirus (HPV)-16 and HPV-18 E6 and E7 genes, in women with progressive cervical cancer. Clinical Cancer Research 2002; 8:3676-3685
• Kurman RJ. Precancerous lesions of the cervix, Chapter 7, Carcinoma and other tumors of the cervix, Chapter 8, in: Blausteirfs Pathology of the Female Genital Tract, Springer Verlag, New York, 1994; 4th ed. pp: 229-299
• Fax H, Wells M, Longocre TA, et al
• Premalignant and malignant squamous lesions of the cervix, Chapter 7, in: Obstetrical and Gynaecological Pathology, Churcill Livingstone, New York, 1995; 4th ed, pp: 273-322
• Karaloğlu D, Tazıcı H, et al. Detection of HPV 16 and HPV 18 infection in patients with cervical neoplasia. Eur J Gynaec Oncol 1996; 17(4): 296-298
• Iwasawa A, Nieminen P, Lehtinen M, et al
• Human papilomavirus DNA in uterine cervix squamous cell carcinoma and adenocarcinoma detected by polymerase chain reaction. Cancer 1996; 77:2275-2279
• Bonfiglio TA, Erozan YS. The biology of human papillomaviruses and their role in cervical carcinogenesis, Chapter 5, in: Gynaecologic Cytopathology. Lippincott-Raven, Philadelphia. 1997; pp: 51-72
• Nakagawa S, Yoshikawa H, et al. Type of human papillomavirus is related to clinical features of cervical carcinoma. Cancer 1996; 78(9): 1935-1941
• Wilczynski SP, Bergen S, et al. Human papillomaviruses and cervical cancer: Analysis of histopathologic features associated with different viral types. Hum Pathol. 1988; 19(6): 697-704
• Huang S, Afonina I, et al. Human papillomaviruses types 52 and 58 prevalent in cervical cancers from Chinese women. Int J Cancer 1997; 70. 408-411
• Bommel P, Brule A, et al. HPV DNA presence and HPV genotypes as prognostic factors in low-stage squamous cell cervical cancer. Gynec Oncol 1993; 48: 333-337
• Chen TM, Chen CA, et al. The genotypes and prognostic significance of human papillomaviruses in cervical cancer. Int J cancer 1994; 57:181-184
• Kraus I, Molden T, Erno LE, et al. Human papillomavirus oncogenic expression in the dysplastic portio; an investigation of biopsies from 190 cervical cones. British Journal of Cancer 2004; 90: 1407-1413
• Matsukura T, Sugase M. Human papillomavirus genomes in squamous cell carcinomas of the uterine cervix. Virology 2004; 324:439-449
• Bachtıary B, Obermair A, Dreier B, et al.Impact of multiple HPV İnfection on response to treatment and survival in patients receiving radical radiotherapy for cervical cancer. Int
• J. Cancer 2002; 102:237-243
• Combita AL, Bravo MM, Touze A, et. al
• Serologic response to human oncogenic papillomavirus types 16,18,31,33,39,58 and 59 virus-like particles in Colombian women with invasive cervical cancer. Int J. Cancer 2002; 97: 796-803
• Aksoy F, Seçkin S. Human papillomavirüs enfeksiyonu ve serviks neoplazileri. Ankara Patoloji bülteni 1992; 9(1): 89-91
• Delvenne P, Fontaine MA, et al. Detection of human papillomaviruses in parafin- embedded biopsies of cervical intraepithelial lesions: Analysis by immunohistochemistry, in situ hybridization and the polymerase chain reaction. Mod Pathol 1994; 7: 113-119
• Konno R, Sato S, Yajima A. Progression of isquamous cell carcinoma of the uterine cervix from cervical intraepithelial neoplasia infected with human papillomavirus. A retrospective follow-up study by in situ hybridization and polymerase chain reaction. Int J Gynec 1992; 11:105-112
• Zehbe I, Wilander E. Human papillomavirus infection and invasive cervical neoplasia: A study of prevalence and morphology. J Pathol 1997; 181:270-275
• Vural G, Tüfekçi C, et al. Türkiye'de yüksek risk içeren bir grupta HPV sıklığı. Jinekoloji ve Obstetrik Dergisi. 1995; 9: 201-204
• Song YS, Kee SH, et al. Major sequence variants in E7 gene of human papillomavirus type 16 from cervical cancerous and noncancerous lesions of Korean women. Gynec Oncol 1997; 66:275-281
• Kristensen GB, Karlsen F, et al. Human papillomavirus has no prognostic significance in cervical carcinoma. Eur J Cancer 1996; 32A(8): 1349-1353
• Rosai J. Female Reproductive System, Chapter 19, inAckermanxs Surgical Pathology, ed Rosai J. St. Louis, Mosby, 2004; 9th ed
• pp:1483-1762
• Einstein BI. The polymerase chain reaction: A new method of using molecular genetics for medical diagnosis. N E J Med 1990; 322: 178-183
• Sasagawa T, Minemoto Y, Basha W, et al
• A New PCR-based assay amplifies the E6-E7 genes of most mucosal human papillomavirus (HPV). Virus Research 2000; 67: 127-139
• Kadish AS, Ho GYF, et al
• Lymphoproliferative responses to human papillomavirus (HPV) type 16 proteins E6 and E7: Outcome of HPV infection and associated neoplasia. J Nat Can Inst 1997; 89(17): 1285- 1292
• Güner H, Barlas MN. Jinekolojik neoplazüerde viral enfeksiyonların yeri ve klinik önemi. Türkiye Klinikleri Jinekoloji Obstetrik Dergisi 1994;.4(2): 67-73
• Koutsky L, Holmes KK, et al. A cohort study of the risk of cervical intraepithelial neoplasia grade 2 or 3 in relation to papillomavirus infection. New Eng J Med 1992; 29:1272-1278
• Dillner J, Lehtinen M, et al. Prospective seroepidemiologic study of human papillomavirus infection as a risk factor for invasive cervical cancer. J Nat Can Inst 1997; 89(17): 1293-1299
• Lorincz A, Reid R, et al. Human papillomavirus infection of the cervix: Relative risk associations of 15 common anogenital types. Obst Gynecol 1992; 79(3): 328-337
• Resnick RM, Cornelissen MTE, et al
• Detection and typing of human papillomavirus in archival cervical cancer specimens by DNA amplification with consensus primers. J Nat Cancer Inst 1990; 82(18): 1477-1484
• Sato S, Takashi O, et al. Sensitivity of koilocytosis, immunohistochemistry and electron microscopy as compared to DNA hybridization in detecting human papillomavirus in cervical and vaginal condyloma and intraepithelial neoplasia. Int J Gynec Pathol 1986; 5(4): 297-307
• Gomez F, Abad MM, Munoz E, et al. Study of infection by human papillomavirus in severe dysplasia and in situ hybridization. Eur J Histochem 1992; 36:137-142
• Schadendorf D, Tiedemann KH, et al.Detection of human papillomavirus in paraffin-embedded condylomata acuminata- comparison of immunohistochemistry, in situ hybridization and polymerase chain reaction
• J Inves Der 1991; 97(3): 549-554
• Basu S, Mitra PK, et al. Detection of human papillomavirus in cervical swabs from Indian women by cytological and immunocytochemical technique. Neoplasma 1991; 38(6): 639-644
• Toki T, Oikawa N, et al. Immunohistoc hemical and electron microscopic demonstra­ tion of human papillomavirus in dysplasia of the uterine cervix. Tohoku J Exp Med 1986; 149:163-167
• Bose S, Choudhuri M, et al
• Immunohisto chemical demonstration of the simultaneous presence of herpes simplex virus type-2 and. human papillomavirus antigens in patients with carcinoma of the uterine cervix
• Med Sci Res 1988; 16: 905-906
• Syrjanen KJ. Human papillomavirus lesions in association with cervical dysplasias and neoplasias.Obst Gynec 1983; 62(5): 617-624
• Reid R, Crum P, et al. Genital warts and cervical cancer. III. Subclinical papillomavirus infection and cervical neoplasia are linked by a spectrum of continuous morphologic and biologic change. Cancer 1984; 943-953
• Benda JA. Pathology of cervical carcinoma and its prognostic implications. Semin Oncol 1994; 21(1): 3-11