Kouichi Fukuda

A comparative analysis of human papillomavirus types 16 and 18 and expression of p53 gene and Ki-67 in cervical, vaginal, and vulvar carcinomas

UNLABELLED This study aimed to investigate the correlation between HPV positivity, p53 overexpression, and cell proliferative activity in cervical, vaginal, and vulvar squamous cell carcinoma. METHODS Sixteen vaginal and 31 vulvar squamous cell carcinomas were examined retrospectively for overexpression of p53 gene and Ki67 antigen by immunohistochemistry and for the presence of HPV types 16 and 18 DNA using a polymerase chain reaction (PCR) method. The results were compared with those obtained from 40 cervical squamous cell carcinomas. RESULTS HPV type 16 or 18 DNA was detected in 21 (52.8%) of 40 cases of cervical carcinomas and p53 overexpression in one (2.5%), while HPV DNA sequences were detected in seven (43.7%) of 16 cases of vaginal carcinoma and p53 overexpression in three (18.7%). With regard to vulvar carcinoma, HPV was harbored in four (12.8%) of 31 cases and p53 overexpression in 19 (61.2%). These results indicated statistically significant inverse correlations between HPV positivity and p53 overexpression (R = -0.999, P < 0.0001). Overexpression of Ki-67 was detected in 28 (70.0%) of 40, 12 (75.0%) of 16, and 21 (67.7%) of 31, cervical, vaginal, and vulvar carcinomas, respectively. There was no significant difference among the three groups. CONCLUSIONS In cervical carcinoma, HPV types 16 and 18 might play a common causal role, and in vulvar carcinoma, p53 gene mutations might be a main causal factor for carcinogenesis. Vaginal carcinoma, on the other hand, is considered to have transitional characteristics between cervical and vulvar carcinoma.

Detection of human papillomavirus genome and analysis of expression of c-myc and Ha-ras oncogenes in invasive cervical carcinomas

Invasive carcinomas of the uterine cervix of 38 patients were examined for the presence of human papillomavirus (HPV) genomes and for the state of the c-myc and Ha-ras oncogenes. A combination of Southern blot hybridization and polymerase chain reaction revealed the presence of the genome of HPV type 16 in 17 tumors (45%), that of HPV type 18 in 3 tumors (8%), and that of unknown types in 16 others (42%), while no viral DNA sequences were detected in 2 tumors. Of the 38 tumors, c-myc amplification was found in only 1 tumor, while there was no Ha-ras amplification. Overexpression of the c-myc gene was observed in 15 (44%) of the 34 tumors analyzed, while there was no overexpression of Ha-ras. Of the 23 squamous cell carcinomas analyzed, relapse-free rates at 24 months were 55% in tumors with c-myc overexpression and 100% in case of tumors with no c-myc overexpression, respectively. The results suggest the possibility that activation of the c-myc oncogene is involved in tumor progression.

Detection of Epstein-Barr virus DNA from a lymphoma-like lesion of the uterine cervix

Abstract The case of a 60-year-old woman in whom a lymphoma-like lesion of the cervix was found during an episode of silent Epstein-Barr virus (EBV) infection is presented. Fractional curettage was performed because of abnormal endometrial smear. The endocervical curettage specimens were diagnosed as highly suggestive of malignant lymphoma, but microscopic examination of a subsequent hysterectomy specimen revealed a benign lymphoid hyperplasia. Those were retrospectively interpreted as a lymphomalike lesion of the cervix. In the absence of clinical symptoms of infectious mononucleosis, the results of serologic tests for EBV revealed an active EBV infection. EBV DNA was demonstrated in nuclei of large lymphoid cells in endocervical curettage specimens by in situ hybridization. She is alive and well 32 months postoperatively. When female patients with lymphoma-like lesions of the lower genital tract are encountered, examinations for EBV are recommended.

Immunohistochemical study of p53 expression in endometrial carcinomas: correlation with markers of proliferating cells and clinicopathologic features

Using anti-p53 (PAb1801 and PAb240), anti-DNA polymerase α and Ki-67 monoclonal antibodies, the expression of p53 was studied in 11 normal endometria, 14 endometrial hyperplasias and 27 endometrial carcinomas and its relationship to the proliferative activity of the tumors was examined. Normal endometria and simple hyperplasias were completely negative for p53. The PAb1801 indices of complex hyperplasias and complex atypical hyperplasias were 2.5±1.8% and 5.0±3.2%, respectively. The PAb1801 indices of grade 1, grade 2 and grade 3 endometrial carcinomas were 10.2±14.2%, 44.4±29/0% and 45.0±32.5%, respectively. These results indicate a progressively enhanced p53 expression in the sequence from normal endometrium, through hyperplasia to carcinoma. A significant correlation between p53 expression and labeling indices of Ki-67 and DNA polymerase α was observed in endometrial carcinomas. The endo-metrial carcinomas with p53 overexpression developed mainly in post-menopausal patients and were frequently high-grade tumors with deep myometrial invasion. These findings may indicate that overexpression of p53 protein contributes to the proliferative activity of the tumor cells.

Prospective follow‐up of Japanese women with cervical intraepithelial neoplasia and various human papillomavirus types

Objective: To evaluate the prevalence of genital human papillomavirus (HPV) types in cervical neoplasias and to evaluate the biological activity of individual HPV types in cervical carcinogenesis. Method: Cellular samples from 318 patients with cervical intraepithelial neoplasia (CIN) or invasive cervical carcinoma were examined for HPV DNA, using a polymerase chain reaction. Of these, 145 women with CIN grade I or II were prospectively followed to better understand the natural history of these precancerous lesions. Result: HPV DNA was detected in 88, 80 and 89%, of CIN grade I, II and III, respectively, and in 92% in invasive carcinomas. The CIN follow‐up data showed a significantly higher progression rate in patients with CIN II than in cases of CIN I, and our classification of HPV types based on HPV prevalence data correlates well with the prospective follow‐up data. A significantly higher progression rate was observed in HPV‐negative CINs. Conclusion: Classification of HPV types according to risk to a malignant state seems to be possible. It seems that HPV negative lesions are likely to progress to a malignant state.

Treatment of advanced cervical cancer by a combination of peplomycin, vincristine, mitomycin-C, and cisplatin

Eighteen patients with advanced or recurrent carcinoma of the cervix were treated with a combination of peplomycin, vincristine, mitomycin-C, and cisplatin (POMP). Ten of the 16 evaluable patients (63%) responded, including 4 with a complete response. Median duration of the response was 7 months. Two of 6 with intrapelvic recurrent tumors responded to some extent following intraarterial infusion. The subcutaneous infusion of peplomycin was well accepted by the patients. Toxicity was tolerable. This regimen seemed to be one of the regimens which should be considered for the advanced or recurrent cervical cancer.

Colposcopic assessment of stage I cervical cancer

A differential assessment by colposcopy of three subgroups of stage I in the new FIGO classification of cervical cancer was attempted. Colposcopic findings in invasive cancer were atypical white epithelium, atypical mosaic, papillary punctation, atypical vessels, abnormal ‘rock-like’ projection, papillomatous growth and ulcer. Ulcer and abnormal ‘rock-like’ projection were more suggestive of a frank invasive carcinoma than a microinvasive carcinoma. While 89% of the cases in stage Ib were correctly predicted, only about 50% of those in stage Ia were correctly predicted. We propose that a colposcopic assessment be included in the armamentarium to decide the method of treatment for cervical cancer patients.

Practical application of the anti-PCNA monoclonal antibody for cytological materials of gynecologic diseases.

Proliferating cell nuclear antigen (PCNA) のモノクローナル抗体である19A2, 19A4, PC10の3種類を用い95%エタノール, 100%メタノール, 4%パラホルムアルデヒドにて10分, 3時間, 24時間固定した細胞標本に免疫細胞学的染色を行った.その結果, 100%メタノールで24時間固定したものに19A2を作用させた場合が最もよい染色性が得られた.その染色条件では正常子宮頸部扁平上皮の表層から中層の細胞は染色されず, 軽度から中等度異形成由来の細胞ではコイロサイトーシスも含め核よりもむしろ細胞質に良好な染色性が得られた. 上皮内癌では核の染色強度が高度異形成より増していた. 浸潤癌では核が強く染色された. 正常増殖期子宮内膜では核に陽性所見を認めたが, 分泌期では認めなかった. 子宮内膜癌では核に陽性所見を認めた. 核異常細胞ならびに悪性細胞100個あたりのPCNA陽性率は核が穎粒状に染色された細胞のみを算出すれば軽度および中等度異形成8.5±3.5%, 上皮内癌15.5±8.9%, 浸潤癌35.8±12.2%でありS期の同定には核の染色態度を考慮することが重要と考えられた. しかしその評価は他の増殖細胞マーカーと比較しさらに慎重に検討する必要がある.

Detection of epstein-barr virus DNA from a lymphoma-like lesion of the uterine cervix

The case of a 60-year-old woman in whom a lymphoma-like lesion of the cervix was found during an episode of silent Epstein-Barr virus (EBV) infection is presented. Fractional curettage was performed because of abnormal endometrial smear. The endocervical curettage specimens were diagnosed as highly suggestive of malignant lymphoma, but microscopic examination of a subsequent hysterectomy specimen revealed a benign lymphoid hyperplasia. Those were retrospectively interpreted as a lymphoma-like lesion of the cervix. In the absence of clinical symptoms of infectious mononucleosis, the results of serologic tests for EBV revealed an active EBV infection. EBV DNA was demonstrated in nuclei of large lymphoid cells in endocervical curettage specimens by in situ hybridization. She is alive and well 32 months postoperatively. When female patients with lymphoma-like lesions of the lower genital tract are encountered, examinations for EBV are recommended.