Nagai N
Department of Obstetrics and Gynecology, Hiroshima University School of Medicine.
Nihon Sanka Fujinka Gakkai Zasshi. 1990 Aug;42(8):823-33.
Recent molecular biologic studies have shown that some type of human papillomavirus (HPV) is closely associated with uterine cervical cancer. In order to investigate the relationship between HPV DNA and cervical cancer, we studied the carcinogenesis of HPV in cervical cancer and related lesions by dot blot hybridization (D.B.H.), Southern blot hybridization (S.B.H.), in situ hybridization (I.S.H.) and polymerase chain reaction (P.C.R.). We also followed some HPV DNA, mRNA positive or negative cases of cervical dysplasia for more than 16 months prospectively. I. Analysis of HPV DNA HPV DNA were detected in cervical dysplasia and cancer and HPV positive rate increased as the grade of cervical dysplasia became higher by several molecular biologic analysis. Especially, the results of I.S.H. with biotinylated HPV DNA probes revealed that HPV DNA was located in the nuclei of koilocytosis, dysplastic cells and cancer cells. In analysis of HPV type in squamous epithelium, HPV 6/11 was the highest positive rate (21.1%) in mild dysplasia. On the other hand, HPV16 positive rate increased with the grade of dysplasia and 34.9% (15/42) of moderate dysplasia and 51.4% (18/35) of severe dysplasia were positive for HPV 16 DNA, respectively. And N/C ratio in HPV 16 positive cells significantly increased by measurement of micrometer. These findings suggest that HPV 16 is high risk HPV and associated with squamous epithelial neoplasia. About 50% of metaplasia close to cervical neoplasia with HPV DNA was positive for the same type of HPV. In columnar epithelium, several types of HPV DNA were detected in 46.7% (15/32) of cervical adenocarcinoma. Thirty one percent (10/32) of adenocarcinoma, 50% (4/8) of adenosquamous cell carcinoma were positive for HPV 18. This suggests an association between HPV 18 and adenocarcinoma and related lesions. We examined amplified DNA detection of HPV 16 and 18 E7 gene by P.C.R. method. HPV 16 and/or 18 DNA were detected in 25 of 43 cases of cervical scrapes obtained from cervical neoplasia, and we confirmed the P.C.R. is the useful method for the screening and the retrospective investigation of HPV infection. By the immunohistochemical and molecular biologic study, there was no correlation HPV DNA with c-myc product and c-myc gene amplification. II. Analysis of HPV mRNA (E6/E7, L1/L2) The early genes E6/E7 of HPV 16 and 18 were considered as one of the carcinogenic factors of uterine cervix. We investigated the localization of HPV E6/E7 and L1/L2 mRNA in cervical dysplasia and CIS with I.S.H. using antisense and sense biotinylated HPV RNA probes which were made by in vitro transcription.(ABSTRACT TRUNCATED AT 400 WORDS)
近期分子生物学研究表明，某些类型的人乳头瘤病毒（HPV）与子宫颈癌密切相关。为了研究HPV DNA与宫颈癌之间的关系，我们通过斑点杂交（D.B.H.）、Southern印迹杂交（S.B.H.）、原位杂交（I.S.H.）和聚合酶链反应（P.C.R.）研究了HPV在宫颈癌及相关病变中的致癌作用。我们还对一些宫颈发育异常的HPV DNA、mRNA阳性或阴性病例进行了超过16个月的前瞻性随访。I. HPV DNA分析 通过几种分子生物学分析方法，在宫颈发育异常和癌症中检测到了HPV DNA，且随着宫颈发育异常程度的升高，HPV阳性率也增加。特别是，用生物素化HPV DNA探针进行原位杂交的结果显示，HPV DNA位于挖空细胞、发育异常细胞和癌细胞的细胞核中。在鳞状上皮的HPV类型分析中，HPV 6/11在轻度发育异常中的阳性率最高（21.1%）。另一方面，HPV16阳性率随着发育异常程度的升高而增加，中度发育异常中34.9%（15/42）、重度发育异常中51.4%（18/35）的HPV 16 DNA呈阳性。通过微米测量发现，HPV 16阳性细胞中的N/C比值显著增加。这些发现表明，HPV 16是高危型HPV，与鳞状上皮肿瘤形成有关。约50%接近宫颈肿瘤且带有HPV DNA的化生组织对同一类型的HPV呈阳性。在柱状上皮中，46.7%（15/32）的宫颈腺癌中检测到几种类型的HPV DNA。31%（10/32）的腺癌、50%（4/8）的腺鳞癌HPV 18呈阳性。这表明HPV 18与腺癌及相关病变之间存在关联。我们通过P.C.R.方法检测了HPV 16和18 E7基因的扩增DNA。从宫颈肿瘤获取的43例宫颈刮片中，25例检测到HPV 16和/或18 DNA，我们证实P.C.R.是筛查和回顾性研究HPV感染的有效方法。通过免疫组织化学和分子生物学研究，HPV DNA与c-myc产物及c-myc基因扩增之间无相关性。II. HPV mRNA（E6/E7、L1/L2）分析 HPV 16和18的早期基因E6/E7被认为是子宫颈致癌因素之一。我们使用体外转录制备的反义及正义生物素化HPV RNA探针，通过原位杂交研究了HPV E6/E7和L1/L2 mRNA在宫颈发育异常和原位癌中的定位。（摘要截于400字）
