중합효소연쇄반응 (polymerase chain reaction:pcr) 에의한 HPV와 EBV의검출 Table 1. Oligonucleotide primers of PCR for HPV and EBV Nucleotide sequence Product size Huma

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1 KISEP Head and Neck Korean J Otolaryngol 2001;44:0611 상악암에서중합효소연쇄반응을이용한인형유두종바이러스와 EpsteinBarr Virus 의검출 조재식 임상철 정형수 임회정 김정현 The Detection of Human Papillomavirus and EpsteinBarr Virus DNA in Maxillary Sinus Carcinoma by Polymerase Chain Reaction Jae Shik Cho, MD, Sang Chul Lim, MD, Hyeong Soo Jeong, MD, Hoi Jeung Lim, MD and Jeong Hyun Kim, MD Department of OtolaryngologyHead and Neck Surgery, College of Medicine, Chonnam National University, Kwangju, Korea ABSTRACT ObjectivesMaxillary sinus carcinoma is rare when compared with cancers of other sites, and its etiology remains unknown. Recent reports demonstrate the possible etiologic role of human papillomavirus HPV and EpsteinBarr virus EBV in maxillary sinus carcinoma. The aim of this study is to detect HPV and EBV in the maxillary sinus carcinoma and examine the relationship between HPV, EBV and maxillary sinus carcinoma. We also compared the clinical features of patients with HPVpositive and HPVnegative to determine the clinical significance of HPV. Materials and MethodThe authors retrospectively searched for HPV and EBV in 40 cases of maxillary sinus carcinoma by using the polymerase chain reaction PCR on DNA extracted from formalinfixed, paraffinembedded tissues. ResultsHPV was detected in of the 40 cases 12.% of maxillary sinus carcinoma, whose histological type was all squamous cell carcinoma. EBV was not detected. Among the five HPVpositives, three were HPV subtype 16 and two were not determined. ConclusionHPV may play a role in the pathogenesis of maxillary sinus carninoma, but EBV needs further study. However, the presence of HPV is not related to Tcategory, cervical metastases, or local recurrence. Korean J Otolaryngol 2001;44:0611 KEY WORDSMaxillary sinus carcinoma PCR Human papillomavirus EpsteinBarr virus. 06

2 중합효소연쇄반응 (polymerase chain reaction:pcr) 에의한 HPV와 EBV의검출 Table 1. Oligonucleotide primers of PCR for HPV and EBV Nucleotide sequence Product size Human papillomavirus HPV consensus primers MY09 CGT CCM ARR GGA WAC TGA TC 40 bp MY11 GCM CAG GGW CAT AAY AAT GG HPV type specific primers 161 ACA GTT ACT GCG ACG TGA GG 239 bp 162 TTT GTT CAG GAC ACA GTG GC 181 TAT ACC GCA TGC TGC ATG CC 7 bp 82 ACG GTT TCT GGC ACC GCA GG EpsteinBarr Virus EBV BV1 TTT TGA ATA CCA CCA AGA AG 82 bp BV2 AAT TTC GGG TTG GAT CCT CC Φ 07

상악암에서 HPV와 EBV의 검출 (Fig. 1), HPV16아형을 검출하기위하여 oligoprimer을 and 4). 또한 상악암조직이 아닌 염증성 용종의 대조군에서 이용한 PCR을 시행한 결과 HPV consensus primer에 양 는 HPV와 EBV가 모두 음성반응을 보였다.

7%)을 나타냈고 선암종과 선양낭성 암종에서는 음성을 나타내었으며, 아형검사에서는 예중 3 Fig. 1. Amplification of a 40bp DNA fragment by PCR using HPV consensus primer. Twelve ul of each reaction mixture is run on 1.

lane 1, 2, 4, 6 and 9 are positive for HPV. Fig. 3. There are no positives in HPV18 subtype among HPV consensus primer positives. Fig. 2. Amplification of a 239bp DNA fragment by PCR using HPV type 16 primer.

lane M, ΦX174 RF/Hae III use as a size marker lane P, positive control lane N, negative control positive ELIZA for HPV type 16/18 in cervical cancer lane 1 maxillary carcinoma of positive PCR for HPV

3 상악암에서 HPV와 EBV의 검출 (Fig. 1), HPV16아형을 검출하기위하여 oligoprimer을 and 4). 또한 상악암조직이 아닌 염증성 용종의 대조군에서 이용한 PCR을 시행한 결과 HPV consensus primer에 양 는 HPV와 EBV가 모두 음성반응을 보였다. 성을 보였던 예중 3예에서 239bp HPV 16형 유전자형 상악암에서 HPV는 총 40예중에서 예(12.%)에서 양 의 band를 확인하였으며(Fig. 2), HPV 18형은 검출되지 않았 성으로 나타났으며, 조직학적으로는 편평세포암종 34예중 고, EBV는 모든 조직에서 음성반응을 나타내었다(Figs. 3 에서만 모두 양성(14.7%)을 나타냈고 선암종과 선양낭성 암종에서는 음성을 나타내었으며, 아형검사에서는 예중 3 Fig. 1. Amplification of a 40bp DNA fragment by PCR using HPV consensus primer. Twelve ul of each reaction mixture is run on 1.9% agarose gel at 100V for 40 min. lane M, ΦX174 RF/Hae III use as a size marker lane P, positive control positive ELIZA for HPV type 16/18 in cervical cancer lane 110 maxillary carcinoma. lane 1, 2, 4, 6 and 9 are positive for HPV. Fig. 3. There are no positives in HPV18 subtype among HPV consensus primer positives. Fig. 2. Amplification of a 239bp DNA fragment by PCR using HPV type 16 primer. Twelve ul of each reaction mixture is run on 1.9% agarose gel at 100V for 40 min. lane M, ΦX174 RF/Hae III use as a size marker lane P, positive control lane N, negative control positive ELIZA for HPV type 16/18 in cervical cancer lane 1 maxillary carcinoma of positive PCR for HPV consensus primer. Lane 1, 3 and 4 are positive for HPV tybe 16. Fig. 4. There are all negative for EBV. Table 2. Clinopathologic data of HPV*positive maxillary sinus carcinoma patients Patients No. Age (yr) Sex 2 64 Male Female Male T Male Female T * human papillomavirus Tcategory Lymph node metastasis Local recurrence HPV* type Table 3. Clinical correlations among HPV, Tstage, cervical lymph node metastases and local recurrence Tstage T2 T3 Cervical nodes Total Positive Negative Local recurrence Total Positive Negative Total HPVpositive HPVnegative Korean J Otolaryngol 2001 ;44 :0611

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5 REFERENCES 1) Gi Min YG, Kang JW, Paik SI, Lee JG, Kwon SH, Suh SH, et al. Cancer of nasal cavity and paranasal sinuses in Clinical Rhinology. The Korean Rhinologic Society 1997: ) Shankar Giri. Cancers of the Maxillary Sinus. Southern Medical Journal 1991;84: ) Syrjanen KJ, Pyrhonen S, Syrjnen SM. Evidence suggesting human papillomavirus(hpv) etiology for the squamous cell papilloma of the parana sal sinus. Arch Geschwulstforsch 1983;3: ) Furuta Y, Takasu T, Asai T, Shinohara T, Sawa H, Nagashima K, et al. Detection of human papillomavirus DNA in carcinomas of the nasal cavities and paranasal sinuses by polymerase chain reaction. Cancer 1992;69:337. ) Clayman GL, Stewart MG, Weber RS. Human papillomavirus in laryn geal and hypopharyngeal carcinomas. Relationship to survival. Arch Oto laryngol Head Neck Surg. 1994;120: ) Kashima HK, Kessis T, Hruban RH, Wu TC, Zinreich SJ, Shah KV. Human papillomavirus in sinonasal papilloma and squmaous cell carci noma. Laryngoscope 1992;102: ) Caruana SM, Zwiebel N, Cocker R, McCormick SA, Eberle RC, Lazarus P. p3 alteration and human papilloma virus infection in paranasal sinus cancer. Cancer 1997;79: ) Lopategui JR, Gaffey MJ, Frerson HF Jr, Chan JK, Mills SE, Chang KL, et al. Detection of EpsteinBarr virus RNA in sinonasal undifferentiated carcinoma from Western and Asian patients. Am J Surg Pathol 1994;18: ) Abramson AL, Brandsma J, Steinberg B, Winkler B. Verruous carci noma of the larynx. Arch Otolaryngol 198;111: ) Brandsma JL, Abramson AL. Association of papillomavirus with cancers of the head and neck. Arch Otolaryngol Head Neck Surg 1989;11: ) Kiyabu MT, Shibata D, Arnheim N, Martin WJ, Fitzgibbons PL. Detection of human papillomavirus in formalinfixed, invasive sq 10 Korean J Otolaryngol 2001;44:0611

6 uamous carcinomas using the polymerase chain reaction. Am J Surg Pathol 1989;13: ) Hoshikawa T, Nakajima T, Uhara H. Detection of human papilloma virus DNA in laryngeal squmaous cell carcinomas by polymerase chain reaction. Laryngoscope 1990;100: ) Syrjanen S, Happonen RP, Virolainen E, Silvonen L, Syrjanen K. Detection of human papillomavirus (HPV) structure antigens and DNA types in inverted papillomavirus and squamous cell carcinomas of the nasal cavities ad paranasal sinuses. Acta Otolaryngol (Stockh) 1987;104: ) Ishibashi T, Matsushima S, Tasnokawa Y. Human papillomavirus DNA in squamous cell carcinoma of the upper aerodigestive tract. Arch Otolar yngol Head Neck Surg 1990;116: ) Howley PM. Role of the human papillomaviruses in human cancer. Cancer Res 1991;1: ) Koutsky LA, Galloway DA, Holmes KK. Epidemiology of genital human papillomavirus infection. Epidemiol Rev 1989;10: ) Macdonald MR, Le KT, Freeman J, Hui MF, Cheung RK, Dosch HM. A majority of inverted sinonasal papillomas carries Epstein Barr virus genomes. Cancer 199;7: ) Gallo O, Di Lollo S, Graziani P, Gallina E, Baroni G. Detection of EpsteinBarr virus genome in sinonasal undifferentiated carcinoma by use of in situ hybridization. Otolaryngol Head Neck Surg 199; 112: ) Leung SY, Yuen ST, Chung LP, Kwong WK, Wong MP, Chan SY. EpsteinBarr virus is present in a wide histological spectrum of sinonasal carcinomas. Am J Surg Pathol 199;19: ) Gaffey MJ, Frieson HF, Weiss LM, Barber CM, Barber GB, Stoler MH. Human papillomavirus and EpsteinBarr virus in sinonasal Schneiderian papillomas. An in situ hybridization and polymerase chain reaction study. Am J Clin Pathol 1996;106:

DNA의추출 Table 1. Sequences of oligonucleotide primers Primers L1 consensus primer Sequence CGT CCM ARR GGA WAC TGA TC Size of amplified products

DNA의추출 Table 1. Sequences of oligonucleotide primers Primers L1 consensus primer Sequence CGT CCM ARR GGA WAC TGA TC Size of amplified products KISEP Rhinology Korean J Otolaryngol 2000;43:836-43 비 부비동반전성유두종에서인유두종바이러스및 Epstein-Barr 바이러스의검출과 p53 단백및 Proliferating Cell Nuclear Antigen 의발현 조진희 서병도 전범조 장한성 양명재 윤희로 노우영 Detection of Human Papillomavirus