자궁경부 상피내 종양 및 침윤성 자궁경부암과 Human papillomavirus type 16 및 type 18 DNA의 상호 연관성에 관한 연구

Abstract

[한글]

Human papillomavirus (HPV) type 16과 type 18은 자궁경부암 및 진행성 경향이 높은자궁경부 상피내종양 조직에서 주로 숙주 세포 염색체내에 삽입된 상태로 높은 출현 빈도를 보이고 있으며(Durst등, 1983 ; Crum등, 1984, 1985 ; Syrjanen등, 1985), 이들 바이러스 genome중에서 특히 transacting factor (Schwarz등, 1985) 또는 promoter/enhancer로 작용 하는 부위(Durst등, 1987) 및 이들 부위의 세포 변형 능력에 대한 연구가 진행중이다(Tsunokawa등, 1986a).

본 연구에서는 HPV type 16, type 18 DNA와 자궁경부 상피내종양 및 침윤성 자궁경부암과의 연관성에 대해 알아보고자 하였다.

자궁경부 상피내종양 12예, 침윤성 자궁경부암 34예 자궁경부의 전이성 선암 1예 및 대조군(만성자궁경부염) 24예 등 총 71예에서 자궁경부 조직을 생검하여 세포내 DNA를 추출한 다음 (32)**P-labelled HPV type 16, type 18 DNA를 각각 probe로 사용하여 dot blotting hybridization 및 자가방사법(autoradiography)을 시행하여 자궁경부 조직에서 HPV type 16 또는 type 18 DNA의 출현 빈도를 조사한 결과 type 16/1ype 18 DNA의 출현 빈도는 침윤성 자궁경부암과 자궁경부 상피내종양군에서 각각 58.8%, 58.3%로서 대조군의 8.4%에 비해 의의있게 높았다. 자궁경부암 환자에서 병기(stage)와 자궁경부 조직의 HPV DNA출현 빈도에는 의의있는 상관 관계가 없었으며, 조직학적 세포 형태별로 볼때 편평상피암에서 HPV type 16/type 18의 출현 빈도는 56.7%였고, 선암 2예 중 1예 및 선편평상피암 1예에서는 type 16과 type 18, type 18이 각각 검출되었다.

자궁경부암 환자의 자궁경부 조직에서 dot blotting hybridization 결과 HPV type 16,type 18DNA가 단독으로 검출되었던 환자중 8예, 5예의 DNA를 각각 여러가지 제한 효소로처리하여 Southern blotting hybridization및 자가방사법을 시행하여 세포내에서 HPV type 16, type 18 genome의 존재 형태를 분석한 결과 HPV type 16 genome은 8예중 7예에서 multiple copy로 숙주세포 염색체내의 여러 부위에 삽입되어 있었고 그 삽입 양상은 다양하였지만 8예에서 모두 바이러스 genome중 PstI으로 절단되어 나타나는 1.78 kb에 해당하는 B조각이 보존되어 있었다. HPV type 18 genome은 5예에서 모두 multiple copy로 숙주세포 염색체내의 여러 부위에 다양하게 삽입되어 있었다.

이상의 연구결과를 볼때, 한국여성에서 자궁경부조직의 HPV type 16, type 18의 감염은 자궁경부의 편평상피암 뿐 아니라, 선암, 선편평상피암의 발생에도 관여할 것으로 사료되며, type 16의 경우 자궁경부 조직 세포내에서 바이러스 genome중 1.78 kb의 B조각의

존재유무가 암의 발생, 진행등에 중요할 것으로 사료된다. 따라서 HPV type 16, type 18DNA가 자궁경부암의 확실한 발암 인자임을 밝히기 위하여 앞으로 이들 바이러스 genome내에서 발암 능력을 갖는 부위를 규명하기 위한 연구가 필요할 것으로 사료된다.

Human Papillomavirus Type 16 and 18 DNAs in Relation to Cervical Intraepithelial

Neoplasia and Invasive Cervical Carcinoma

Dong Hee Choi

Deparment of Medical Science The Graduate School, Yonsei University

(Directed by Professor Tchan Kyu Park, M.D., Ph.D.)

Human papillomavirus (HPV) type 16 and 18 DNAs are known to be found frequently

in the biopsy specimens from the lesions of both cervical intraepithelial neoplasia

(CIN) with high tendency for progression and invasive cervical carcinoma (ICC), and

their integration into human genome has been documented (Durst et al,1983; Crum et

al,1984; Syrjanen et al,1985). It has been suggested that some regions within the

viral genome may play some role as a transacting factor (Schwarz et at, 1985) or a

promoter/enhancer (Durst et al, 1987, and efforts are now directed to showing the

transforming activity of these regions (Tsunokawa et al, 1986a).

The purpose of this study is to identify the relationship of HPV type 16 and 18

DNAs to CIN and ICC by analyzing the occurrence of their DNA sequences in the cells

of the cervical tissues and their physical states in the cancerous cervical

tissues.

Cellular DNAs were extracted from the cervical tissues of a total of 71 cases; 21

CINs, 34 ICCs, 1 metastatic adenocarcinoma of the cervix, and 24 controls with

chronic cervicitis. Using (32)**P-labelled HPV type 16 or 18 DNA as a probe, HPV

DNA typing was performed by dot blotting hybridization and autoradiography. The

following results were obtained: The positive rates for HPV type 16 and/or 18 DNA

in the ICC and CIN cases were 58.8% and 58.3% respectively; they were significantly

higher than that of the control group (8.4%). In the cases with ICC, no significant

difference in positive rate for HPV DNA according to stage was noted. According to

histological cell type, HPV type 16 and 18 DNAs were detected in 43.3% and 23.3%

each in squamous cell carcinoma cases. HPV type 16 and 18 DNAs were detected in 1

of 2 cases of adenocarcinoma, and only HPV type 18 DNA was detected in 1

adenosquamous carcinoma case.

Cellular DNAs from 8 and 5 cases of ICC each positive in HPV type 16 DNA alone

and type 18 DNA alone were treated with several restriction enzymes. They were then

processed with Southern blotting hybridization and autoradiography to evaluate the

physical states of their viral genomes. Seven of the 8 cases haying HPV type 16

genome and all 5 cases having HPV type 18 genome had their viral genomes integrated

as multiple copies at multiple sites of their host chromosomes, and the patterns of

integration varied. It was noted that B band of 1.78kb of HPV type 16 genome was

retained in all 8 cases.

This study demonstrated that HPV type 16 and 15 DNAs in cervical tissues might be

considered to be potentially important in the development of cervical cancer of

various cell types including adeno-, adenosquamous as well as squamous cell

carcinomas. And the role of HPV type 16 in He development or progression of

cervical cancer in the presence of 1.78kb of B fragment of its viral genome in

cervical tissue was suggested. It seems that further research of several specific

sites having oncogenic potential within the viral genome would be desirable to

determine the causative nature of HPV type 16 and 18 DNAs in cervical carcinoma.

[영문]

Human papillomavirus (HPV) type 16 and 18 DNAs are known to be found frequently in the biopsy specimens from the lesions of both cervical intraepithelial neoplasia (CIN) with high tendency for progression and invasive cervical carcinoma (ICC), and their integration into human genome has been documented (Durst et al,1983; Crum et al,1984; Syrjanen et al,1985). It has been suggested that some regions within the viral genome may play some role as a transacting factor (Schwarz et at, 1985) or a promoter/enhancer (Durst et al, 1987, and efforts are now directed to showing the transforming activity of these regions (Tsunokawa et al, 1986a).

The purpose of this study is to identify the relationship of HPV type 16 and 18 DNAs to CIN and ICC by analyzing the occurrence of their DNA sequences in the cells of the cervical tissues and their physical states in the cancerous cervical

tissues.

Cellular DNAs were extracted from the cervical tissues of a total of 71 cases; 21 CINs, 34 ICCs, 1 metastatic adenocarcinoma of the cervix, and 24 controls with chronic cervicitis. Using (32)**P-labelled HPV type 16 or 18 DNA as a probe, HPV DNA typing was performed by dot blotting hybridization and autoradiography. The following results were obtained: The positive rates for HPV type 16 and/or 18 DNA in the ICC and CIN cases were 58.8% and 58.3% respectively; they were significantly higher than that of the control group (8.4%). In the cases with ICC, no significant

difference in positive rate for HPV DNA according to stage was noted. According to histological cell type, HPV type 16 and 18 DNAs were detected in 43.3% and 23.3% each in squamous cell carcinoma cases. HPV type 16 and 18 DNAs were detected in 1

of 2 cases of adenocarcinoma, and only HPV type 18 DNA was detected in 1 adenosquamous carcinoma case.

Cellular DNAs from 8 and 5 cases of ICC each positive in HPV type 16 DNA alone and type 18 DNA alone were treated with several restriction enzymes. They were then processed with Southern blotting hybridization and autoradiography to evaluate the

physical states of their viral genomes. Seven of the 8 cases haying HPV type 16 genome and all 5 cases having HPV type 18 genome had their viral genomes integrated as multiple copies at multiple sites of their host chromosomes, and the patterns of

integration varied. It was noted that B band of 1.78kb of HPV type 16 genome was retained in all 8 cases.

This study demonstrated that HPV type 16 and 15 DNAs in cervical tissues might be considered to be potentially important in the development of cervical cancer of various cell types including adeno-, adenosquamous as well as squamous cell

carcinomas. And the role of HPV type 16 in He development or progression of cervical cancer in the presence of 1.78kb of B fragment of its viral genome in cervical tissue was suggested. It seems that further research of several specific sites having oncogenic potential within the viral genome would be desirable to

determine the causative nature of HPV type 16 and 18 DNAs in cervical carcinoma.