Pyrimidine nucleoside phosphorylase activity in normal tissues of the uterus and ovary and in benign and malignant lesions of these organs.

BACKGROUND: Pyrimidine nucleoside phosphorylase (PyNPase) is identical to the protein, platelet-derived endothelial cell growth factor (PD-ECGF), which has angiogenic activity. The physiological roles of PyNPase activity in the uterus and ovary are not known. In this study, we measured PyNPase activity in normal tissues of the uterus, ovary, and lymph nodes, and in benign and malignant lesions of these organs, and we considered the clinical implications of PyNPase activity in the uterus and ovary. METHODS: Tissue samples were obtained from 163 patients (whose diseases are listed below) during surgery. PyNPase activity was measured spectrophotometrically, by monitoring the formation of 5-fluorouridine. RESULTS: Mean PyNPase activity in tissues from the lesions of patients with cervical cancer (n = 20), uterine endometrial cancer (n = 26), leiomyoma (n = 23), ovarian cancer (n = 46), ovarian endometriosis (n = 21), and benign epithelial ovarian tumor (n = 27) was significantly greater than that in the corresponding normal tissues. The PyNPase activity in the normal endometrium was significantly higher in the secretory phase than in the proliferative phase. The activity in normal or metastatic lymph nodes was significantly greater than that in normal tissues of the uterus and ovary. Mean PyNPase activity in cancerous cervical tissues was significantly greater than that in cancerous endometrial tissues or cancerous ovarian tissues. There were no significant differences in PyNPase activity in cervical cancer, endometrial cancer, and ovarian cancer tissues according to tumor stage. The enzyme activity appeared to be greater in histopathological G3 grade endometrial cancer than in G1 and G2 endometrial cancer. The enzyme activity in mucinous adenocarcinoma of the ovary was significantly lower than that in serous, endometrioid, and clear cell adenocarcinomas. All patients with cervical squamous cell carcinomas with PyNPase activity greater than 500 nmol/min per mg protein exhibited lymph node metastasis. CONCLUSION: Increased PyNPase activity consistently reflected neoplastic growth, and varying levels of activity were seen in different histologic cell types. This enzyme activity may be involved in cervical squamous cell carcinomas, in normal and metastatic lymph nodes, and in the normal, secretory phase in the endometrium.

Sodium butyrate induces growth arrest and senescence-like phenotypes in gynecologic cancer cells.

We demonstrated here the growth-suppressing effects of sodium butyrate (NaB) on human endometrial and ovarian cancer cells. The arrest of cells at the G1 checkpoint accounted for this effect. NaB-mediated p21 might arrest endometrial and ovarian cancer cells at the G0/G1 phase by eliciting pRb unphosphorylation. To demonstrate the role of pRb regulation by p21, we measured the sensitivity to NaB of cervical cancer cells in which pRb had been inactivated by HPV E7. The cervical cancer cells displayed a sensitivity in NaB-mediated G2/M arrest in addition to their sensitivity in G0/G1 arrest. Arrest at G0/G1 and G2/M accompanied induction of senescence-like phenotypes (SLPs). Most importantly, the effect of NaB on senescence induction was not coupled with the predominance of hypophosphorylated pRb forms in the cervical cancer cells. This suggested that NaB had the potential to elicit SLPs through p21-mediated withdrawal from cell cycle progression. The consequences of p21 induction were manifold. The effects of NaB on gynecologic cancer cell growth indicated its potential use in cancer treatment. NaB was effective even in the cancer cells with mutant p53 and/or Rb genes by eliciting cell senescence.

New method to prevent bladder dysfunction after radical hysterectomy for uterine cervical cancer.

OBJECTIVE: The purpose was to improve the surgical procedures to prevent bladder dysfunction after radical hysterectomy. METHODS: Twelve patients with stage Ib cervical cancer underwent intraoperative electrical stimulation to identify the vesical branches of the pelvic nerves. Autonomic nerve localization in the vesicouterine ligament was examined in 10 patients immunohistochemically. According to the results of the above studies a new method to preserve the vesical branches was developed. Grades of postoperative bladder dysfunction were compared between new (n = 19) and conventional methods (n = 18). RESULTS: Electrical stimulation on the outer surface of the posterior sheath of the vesicouterine ligament caused the increase of intravesical pressure. S-100 protein localized also on this area. Postoperative compliance of the detrusor in cases with the new method demonstrated less decrement from preoperative values than in cases with the conventional method. The new method required significantly fewer days to achieve residual urine volumes less than 50 ml after surgery. CONCLUSIONS: The new method significantly reduces bladder dysfunction after radical hysterectomy.

[Clinical results of repeated intraperitoneal administration of CDDP in patients with ovarian cancer].

CDDP was intraperitoneally administered to 45 ovarian cancer patients (9 cases in Stage Ic, 9 cases in Stage II, 19 cases in Stage III and 8 cases in Stage IV) as the first line chemotherapy to examine response rates and cumulative survival rates by Kaplan-Meier method. Among 18 cases with measurable tumor, there were 5 CR cases, 6 PR cases, 4 NC cases and 3 PD cases. Eleven out of 18 responded, for a response rate of 61.1%. The 3- and 5-year survival rates were 88.9% and 88.9% in Stage I, 46.7% and 0% in Stage II, 48.6% and 41.7% in Stage III, and 0% and 0% in Stage IV, respectively. The 3- and 5-year survival rates in patients with residual tumor smaller than 2 cm in diameter after the first surgery in Stage III or IV were 76.9% and 61.4% respectively, in contrast to 11.5% and 11.5% in patients with residual tumor of 2 cm or larger in diameter. Thus, a significantly better prognosis was demonstrated in patients with residual tumor smaller than 2 cm. These results indicate that intraperitoneal administration of CDDP demonstrated high response rates and proved to be useful as the first line chemotherapy, yet did not manage to improve the long-term prognosis of patients with progressive cancer.

[Pharmacokinetics of CDDP by the route of administration in patients with ovarian cancer].

CDDP was administered to 36 patients with ovarian cancer by one of three routes: i.v., ip, or ia, and its pharmacokinetics was studied. 1) With an initial dose of 100 mg/body the peak values for blood free-pt were 1.63 +/- 0.78 micrograms/ml (S.D.) in the i.v. group, 0.61 +/- 0.53 micrograms/ml (S.D.) in the ip group, and 1.62 +/- 0.78 micrograms/ml (S.D.) in the ia group. The area under the curve (AUC) was 2.56 +/- 0.72 micrograms.hr/ml (S.D.), 1.84 +/- 0.75 micrograms.hr/ml(S.D.), and 2.70 +/- 0.51 micrograms.hr/ml (S.D.), respectively. 2) The free-pt level of the ascitic fluid in the ip group was 25.3 +/- 28.1 micrograms/ml (S.D.) just after the injection of 100 mg/body. 3) The ovarian tissue level following the initial CDDP dose of 100 mg/body was the highest in the ia group, next highest in the ip group, and lowest in the i.v. group. In some cases, the ovarian tissue level was markedly elevated after repeated ip injections. Deep-middle layer concentrations in ovarian tissue were not so low in the ip group as in the i.v. or ia group. As noted above, the blood free-pt peak was low following ip administration, but the ovarian tissue level was higher following ip administration than following i.v. though lower than following ia administration. These findings suggest that ip administration of CDDP may be useful in treating large tumor masses. Good results are expected particularly following repeated ip therapy, which sometimes provided a markedly high tissue level.

[Sequential changes of HPV DNA, the expression of EGF-R and CD-14, and histopathological findings in HPV infected benign epithelium and dysplastic lesions of the uterine cervix].

In regard to cases in which HPV DNA was detected in the uterine cervix by the Southern blot method, we studied both changes in HPV DNA and histological diagnosis, and moreover we studied the relationship between CD-14 (monocyte/macrophage), EGF-R, and HPV DNA. 1) Changes in HPV DNA, regardless of the HPV type in benign lesions of the uterine cervix, almost all became negative within 1 year. 2) In 1 year benign lesions, the manifestation frequency of CD-14 was higher from the initial examination and during the subsequent 3 months when HPV DNA was detected. 3) HPV DNA in cases of mild dysplasia was detected after 1 year in 41% (7/17), but it continued for 3 years only in 3 cases. 4) Cases of moderate to severe dysplasia, which are though to have developed cancer at an early period of within 1-1.5 years included 26.7% (4/15), and HPV DNA was continually detected in them. The characteristics of these cases were: (1) The HPV 16 type was the main type. (2) The mean age was 34.8 years, which was younger than in regressed cases with a mean age of 43.1 years. (3) The dysplasia occupation rate in the uterine cervix was higher. 5) In 7 cases of moderate to severe dysplasia which regressed from squamous metaplasia, HPV DNA was detected at 1 year in 71.4% (5/7), but at 2-3 years it became undetectable in all cases, so that it has a close relationship with this histological regression.(ABSTRACT TRUNCATED AT 250 WORDS)

[Production of CA125 in cell lines derived from human ovarian carcinoma: in relation to the cell cycle].

The association of the production of CA125 with the cell cycle was investigated in two cell lines derived from human ovarian cancer, one from a serous cystadenocarcinoma (HTOA) and the other from a mucinous cystadenocarcinoma (RMUG-s). HTOA and RMUG-s cells secreted CA125 at about 50 and 30U/ml/10(5) cell/24hr, respectively, in the logarithmic growth phase and at about 75 and 100U/ml/10(5) cell/24hr in the steady phase. Analysis by FCM revealed that cultures of both cell lines cultured for 7 days contained more cells in the G0/G1 phase and less cells in the S phase than those cultured for 3 days. The positive rate of immunologically stained DNA polymerase alpha was 31% in HTOA cells and 39% in RMUG-s cells after cultivation of the cells for 3 days. The addition of EGF at 0.01, 0.1 or 1.0nM did not affect the production of CA125 in HTOA or RMUG-s cells while the addition of NaBT at 1, 3 and 5mM raised production in both cell lines as the dose rose. With RMUG-s cells, the addition of EGF at 0.01nM to the culture media accelerated both logarithmic and steady phase growth without a significant change in the production of CA125. In contrast, the addition of NaBT at 1mM suppressed growth, but tended to increase the production of CA125 per cell. With the effect of EGF on the cell cycle of both cell lines, cells in the S phase increased by about 20% as compared with the control, 48 hours after its addition at 0.01nM. In contrast, after cultivation for 48 hours in the presence of 1mM NaBT, cells in the S phase were decreased while those in the G0/G1 phase increased. The results presented above suggested the possibility that some factors other than the cell cycle were involved in the production of CA125. There also is close correlation between cells in the G0/G1 phase and the production of CA125 in the culture of human ovarian cancer cells.

[Immunohistochemical studies on epidermal growth factor receptor (EGF-R) in gynecological malignant tumor].

The expression of Epidermal Growth Factor Receptor (EGF-R) in gynecological malignant tumors was investigated immunohistochemically. 1) With respect to the expression of EGF-R in the uterine cervix, it was seen in 20.0% with benign lesion. In cases of dysplasia, it was expressed in 62.5% of the cases with mild dysplasia, 81.8% with moderate dysplasia and 53.3% with severe dysplasia. In cases of CIS, it was seen in 46.7% and in cases of invasive cancer, it was seen in 22.2%. By hystological type, the expression rates were 27.3% for keratinized squamous cell carcinoma and 33.3% for large cell non-keratinized squamous cell carcinoma. No expression was seen in three cases of small cell non-keratinized squamous cell carcinoma or four cases of adenocarcinoma. In cases of benign lesions, EGF-R was localized in the cell walls of the basal layer, but in cases with dysplasia, it was found in the cell walls and also in the cytoplasm in all layers of the epithelium. 2) The expression rate in endometrial carcinoma was 14.3% and all of these cases were well-differentiated adenocarcinoma. There was no reverse correlation with estrogen receptors. 3) The expression rate for advanced malignant ovarian tumors was 31.4% and there was no clear correlation with the histological type. The prognosis tended to be better in cases expressing EGF-R than in those not. These results indicated that EGF-R appears to be related to the degree of advance of cervical dysplasia, but it was clear that the frequency of expression of EGF-R decreased when the cancer became invasive. In cases of malignant ovarian tumors, the expression of EGF-R tended to be related to the prognosis.

[Diagnosis and follow-up study of human papillomavirus infection in benign epithelium and mild dysplasia of the uterine cervix].

In this study, we evaluated the sensitivity of cytology, colposcopy and the dot blot method (Vira Pap, Vira Type) for the screening of HPV infection in benign epithelium and mild dysplasia of the uterine cervix, and then investigated subsequent changes in HPV DNA in the lesion. The HPV infection detection rates for cytology, colposcopy and the dot blot method were 14/46 (30.4%), 30/46 (65.2%) and 35/46 (76.1%), respectively. Cases where HPV DNA was negative with the dot blot method but HPV infection was suspected with cytologic and colposcopic examination were 3/6 (50.0%) in the undigestive type except for HPV types 6, 11, 16, 18, 31, 33 and 35. HPV DNA disappeared after 3-6 months in persistent cases with histologically benign epithelium in the one year follow-up. In persistent cases of mild dysplasia followed up histologically for one year from the time of the first visit showed differences between negative and positive cases of HPV DNA after 1 year. In 2 progressive cases (HPV 31 and 16 types) HPV DNA in the cervix was detected continuously. We consider that the dot blot technique is a valuable method for identifying human papillomavirus infection. However, detection of undigestive HPV types by the dot blot method was poor. Therefore, the joint use of cytology, colposcopy and the dot blot method seems to be a valuable screening technique. Individual variations in the clinical course of the high risk group appear to reflect profound differences in the effectiveness of host responses. Therefore, cases where continuous HPV DNA was observed in the high risk groups should be followed up for a long period.