Clinical application of subrenal capsule assay in ovarian cancer

The 6-day subrenal capsule assay (SRC) in normal immuno-competent mice was used to test the responsiveness of ovarian cancer to combination chemotherapy and assess the usefulness of SRC in clinical application. A total of 18 different patients in 22 different assays were studied. Twenty-one assays (95%) were evaluable. The predictive sensitivity was 50%, the predictive resistance 75% and the predictive total accuracy 55% respectively. Sensitive drugs in repeated assays in four patients were not changed. Etoposide, cyclophosphamide, cis-platin and adriamycin were sensitive. Etoposide was the most sensitive of four drugs. The response rate was 50% in the patients treated with etoposide as the first line chemotherapy and the second line chemotherapy, respectively. Patients who were treated with sensitive drugs in SRC survived longer than the patients treated with cis-platin combined chemotherapy without etoposide. The median survival time was 19 months and the mean progression free interval was 15 months. In the results of this study, the value of SRC was supported by the results of chemosensitivity tests and etoposide seems useful in chemotherapy in ovarian cancer.     

Fundamental studies of subrenal capsule assay

Cancer chemosensitivity test by Bogden's SRCA (subrenal capsule assay) method was experimentally evaluated using DMBA induced rat ovarian cancer. Cancer tissues were implanted beneath the renal capsule of BDF1 mice, and factors affecting development of the xenografts were examined. To observe the effect of the host-mediated immunologic reaction, the mice were put into immunosuppressive situations by applying cyclophosphamide (CPA) or cyclosporin A (CsA), and the growth of the tumor in each condition received macroscopic and histological evaluation. Under the most favorable conditions, drug sensitivity tests were performed. The results obtained were as follows: 1) In non-treated mice, the cancer cells of the grafted tumor were rejected, until the 7th day following the implantation, by host reaction. 2) In a CPA-induced immunosuppressive condition, the tumor tissue was damaged by the cytotoxic effect, although the size of the tumor was on the increase until the 9th day. 3) CsA brought about a favorable condition for tumor development, showing a linear growth curve from the 3rd to the 9th day with viable cancer cells. 4) Chemosensitivity tests indicated that methotrexate and CDDP were effective, while mitomycin was not effective in treating the implanted tumors. 5) The results obtained with SRCA were much the same in experimental therapeutic trials using rats with primary ovarian cancer induced by DMBA application.     

Fundamental studies on the subrenal capsule assay as chemosensitivity test for nonsolid tumors

Fundamental evaluation of the subrenal capsule assay (SRCA) method in nonsolid tumors was made, using two types of murine malignant ascites. Malignant ascites were obtained from mice bearing M-5076 ovarian reticular cell sarcoma or MH-134 hepatoma. These tumor cells were allowed to settle by standing at 4 degrees C to form a jelly-like clot. This clot was cut into fragments about 1mm3 in size and one of these fragments was mashed in trypan blue to estimate the viability grade of the implanted tumor cells. The rest of the fragments were implanted beneath the renal capsule of the mice. On the 6th day after implantation, the assay mice were killed, the increase in the size of the tumor was determined and histological examination was carried out. The results were as follows: (1) The clot was formed reproductively by allowing ascites to settle for one or two days and there was a high viability rate for the tumor cells: 79.9 +/- 11.0% of M-5076 and 90.1 +/- 5.9% of MH-134. (2) The ascites clot thus implanted grew rapidly in the control groups but growth was inhibited by chemotherapy: Tumors were reduced significantly (p less than 0.05-0.005) in the group treated with a single agent. This trend towards a suppressive effect of carcinocidal agents on the tumor growth was more conspicuous as a combination regimen was utilized, a combination of three agents producing the maximum effect. (3) The clot grew more quickly than the solid tumor in both the control and the treated groups. There was a high correlation (r = 0.93 in M-5076, and r = 0.64 in MH-134) between the growth rates of ascites and solid tumor in SRCA. (4) Histological examination revealed that viable tumor cells infiltrated widely under the renal capsule in both types of tumors. These results suggest that ascites and solid tumor are useful materials for the subrenal capsule assay method.     

Concordance of combination and single agent chemosensitivity prediction in ovarian carcinoma using the subrenal capsule xenograft assay (SRCA)

The tumors from 62 patients with advanced ovarian adenocarcinoma were assayed by the subrenal capsule xenograft assay (SRCA) for sensitivity to doxorubicin (A), cis-platinum (P), and cyclophosphamide (C), individually and in combination. In some instances only one or two of the individual drugs were assayed; however, the combination, CAP, was always tested. All patients received an optimal surgical debulking (absence of any residual tumor masses greater than or equal to 2 cm) followed by chemotherapy with CAP. Forty-two tumors were predicted to be sensitive to CAP by the SRCA; 51 of 71 (72%) individually tested drugs agreed with this determination. Twenty-one tumors were predicted to be resistant to CAP and 32 of 36 (89%) individually tested drugs agreed with this determination. In this preliminary study, 11 patients had surgically documented partial responses to CAP chemotherapy. All of these patients had tumors which prospectively tested as sensitive to CAP in the SRCA: 13 of 18 (72%) of separately tested drugs were in concordance with this sensitivity. Fourteen patients failed CAP therapy and three of these failures were predicted prospectively by the SRCA: 9 of 9 (100%) of separately tested drugs were in concordance. Thus, there is an overall concordance of 82% (22/27) between the individual components of a combination chemotherapy and the combination therapy itself. It would seem that extrapolations of sensitivity or resistance can be made from the individual components.     

Responsiveness of gynecologic tumors to chemotherapeutic agents in the 6-day subrenal capsule assay

Various gynecologic malignancies have been tested in the 6-day subrenal capsule assay, which is an in vivo test of human tumor responsiveness to drug therapy. Fresh surgical explants of ovarian, endometrial, and cervical tumors were implanted as 1-mm³ fragments under the renal capsule of normal mice and tested against a spectrum of clinically active agents. Regardless of the site of origin, human tumors showed variations in growth rate when implanted under the renal capsule that appeared to reflect both the growth potential characteristic of each tumor as well as the heterogeneity of the cell populations comprising each tumor. An average of 60% of tumors showed positive growth and 11% demonstrated no measurable change in size. The response rates of 18 ovarian, 28 endometrial, and 20 cervical carcinomas to clinically active chemotherapeutic agents were determined. A range of responses, in terms of drugs indicated to be active and of the degree of responsiveness to active agents, was obtained with each histologic type. Response rates varied from 6% to tamoxifen in cervical carcinomas to 80% to 5-fluorouracil in ovarian carcinomas. The results of this study support the variability in chemotherapy responsiveness observed clinically with gynecologic tumors and suggest the feasibility of using the subrenal capsule assay as a predictive test.     

Localization of gonadotropin binding sites in human ovarian neoplasms

The binding of human luteinizing hormone and human follicle-stimulating hormone to ovarian tumor biopsy specimens from 29 patients was analyzed. The binding sites for human luteinizing hormone were demonstrated in one tumor of epithelial origin (mucinous cystadenoma) and in one of sex cord-stromal origin (theca cell tumor). The binding sites for human follicle-stimulating hormone were found in three tumors of epithelial origin (serous cystadenoma and mucinous cystadenoma) and in two of sex cord-stromal origin (theca cell tumor and theca-granulosa cell tumor). The surface-binding autoradiographic study revealed that the binding sites for gonadotropins were localized in the stromal tissue. The results suggest that gonadotropic hormones may play a role in the growth and differentiation of a certain type of human ovarian neoplasms.     

Epithelial ovarian neoplasms of low malignant potential

This report presents 13 patients with epithelial ovarian neoplasms of low malignant potential. Ten patients had serous tumors, 1 had a mucinous tumor, and 2 had endometrioid tumors. Two had Stage I disease, 4 had Stage II disease, and 7 had Stage III disease. All were treated with melphalan. Second-look laparotomy was performed in 11 patients and 1 patient required subsequent exploratory laparotomy. None achieved histologic confirmation of cure. Five patients continued treatment with melphalan and underwent third-look laparotomy with findings of persistent disease. Seven patients are alive without clinically detectable evidence of disease. Four patients died of their disease and 2 patients died of melphalan-induced acute leukemia.     

Establishment of subrenal capsule xenografts of primary human ovarian tumors in SCID mice: potential models

OBJECTIVE: To evaluate subrenal capsule xenografting of primary ovarian tumor tissues in mice for development of new ovarian cancer models. METHODS: Pieces (1 x 3 x 3 mm) of ovarian tumor specimens from patients were meticulously grafted under renal capsules of female NOD/SCID mice within 2 h of surgical removal. Tumor types included papillary serous adenocarcinomas, borderline and benign mucinous cystadenomas, granulosa cell tumors, a serous borderline tumor and a grade 3 mixed surface epithelial tumor of transitional and undifferentiated types. After 1-2 months, grafts were retrieved for comparison with original tissues. Hematoxylin and eosin (H&E) and immunohistochemical staining was carried out using tissue micro-arrays and CEA, B72.3, WT-1, OC125, keratin, inhibin, CK7, CK20, Cam5.2, and MIB-1 as markers. RESULTS: Tumor tissue engraftment rate was > 95%. Comparison of donor and post-graft tissues showed highly similar histopathological features; 91 +/- 5% concordance in immunostaining indicated major preservation of immunophenotypes in the xenografts for 30-60 days. There was a small, but significant, increase in MIB-1 proliferative index in xenografts compared to original specimens. CONCLUSIONS: Subrenal capsule xenografts of primary human ovarian tumors in SCID mice can retain major histopathological and immunohistochemical characteristics of the original tissues. The achievable, consistently high engraftment rate allows use of such xenografts as tools for studying a wide range of ovarian tumors, including granulosa cell tumors and benign, borderline, and malignant surface epithelial neoplasms. Potential applications include preclinical testing of patients' tumor responses to various chemotherapeutic regimens, evaluation of novel therapeutic agents, analysis of tumor progression at cellular and molecular levels, and identification of new therapeutic targets.     

Evaluation of the subrenal capsule assay (SRCA) in breast cancer using immunocompetent (NIC) mice

To assess the value of subrenal capsule assay (SRCA) for predictive chemo-sensitivity testing in breast cancer 24 primary tumours were examined in normal immunocompetent (NIC; AB- and B6D2-F1-) mice. In 6 cancers the effect of 0.25 mg Prednisolone mous X day (applicated with drinking water) was compared to the control group. Independent of the mice-species a significant increase of the grafts size occurred in 14 of 20 evaluable cancers (p less than 0.05). Histologically the increase of tumour size was caused by infiltrated host cells, only in 22 of 107 (20.6%) grafts vitale tumour cells were detectable. After immunosuppression the increase of the grafts size was significant lower, than in the no treated control group (p less than 0.05). In groups treated with Prednisolone only a thin inflammatory host cell infiltration was observed, but vital tumour cells were only seen in 3 of 29 (10.3%) grafts. Conclusion: Using normal immunocompetent mice subrenal capsule assay is not suited for predicting responsiveness of breast cancers to cytostatic drugs.     

Estimation of medroxyprogesterone acetate against a human endometrial tumor constituted from the established Ishikawa cancer cells by a subrenal capsule assay]

In order to establish a sensitivity test system for the evaluation of anti-cancer hormonal agents, we tried a long-term subrenal capsule (SRC) assay, using nude mice with a transplanted solid tumor of endometrial cancer cells (Ishikawa's line). Unlike DNA-affecting agents, anti-cancer hormonal agents exert cytostatic effects rather than cytocidal effects, and their evaluation in a short period is considered to be inaccurate. Our test system is somewhat difficult in terms of technique, but it is useful since it can (1) evaluate the agents in the relatively short period of 28 days and (2) compare the cytostatic anti-tumor efficacy of two or more agents under the same conditions. The rate of successful tumor transplantation in nude mice in our system is very high, i.e., more than 90%. Although there are some points which need improvement, our system is considered to be useful as an assay system for the development of anti-cancer hormonal agents and other similar chemotherapeutic agents for cancers. When medroxyprogesterone acetate (MPA) was evaluated by means of this system, the administration period, as well as the dosage, was found to be important.     

Determination of epidermal growth factor receptor expression in malignant ovarian tumors using radioligand assay

The objective of the authors was to determine in a prospective study EGFR level of expression in ovarian cancer. For a period of 2 years EGFR expression was studied in 23 patients having malignant ovarian tumors by 125I-labelled human epidermal growth factor and recombinant human epidermal growth factor. EGFR expression was observed in 56.5% of the cases. Insignificant mucinous and clear cell cancers prevailed in the EGFR/+/ group. All patients having wider metastasis were EGFR/+/, as this dependency was statistically significant (p = 0.017). Radioligand binding assays enabled exact quantitative determination of EGFR expression in malignant ovarian tumors. Statistically significant relation existed between the ovarian cancer metastatic potential and EGFR expression level.     

卵巢子宫内膜异位症恶变26例临床病理分析

目的 探讨探讨卵巢子宫内膜异位症（内异症）恶变的临床病理特征。方法 回顾分析26例卵巢内异症恶变患者的临床和病理资料。结果 患者以痛经和检查发现盆腔肿块为主要临床表现。行B超或彩色多普勒超声检查者18例,其中10例发现盆腔肿块中含实质性结构。肿瘤组织类型以内膜样腺癌和透明细胞癌常见。58％（15／26）患者的肿瘤显微镜下可见不典型内异症。协际妇产科联盟分期：Ⅰ期21例（81％）,Ⅱ期3例（12％）,Ⅲ期2例（8％）。结论 卵巢内异症恶变早期临床诊断存在困难,B超或彩色多普勒超声检查有重要参考价值。重视观察异位内膜组织形态变化,有利于认识卵巢内异症恶变的发生和发展过程,提高诊断和治疗水平。     

Responsiveness of gynecological malignancies to oral antitumor agents in subrenal capsule assay and individualization of oral adjuvant chemotherapy

A subrenal capsule assay (SRCA) was performed to test the sensitivity of 45 gynecological malignancies, including 24 cervical and 15 ovarian carcinomas, to oral antitumor agents, UFT, cyclophosphamide (CPM) and carboquone (CQ). Additionally, using a human endometrial carcinoma (Ishikawa carcinoma), the utility of SRCA in oral adjuvant chemotherapy was also investigated. Thirty-six of 45 cases (80.0%) were found to be evaluable. Regardless of the origin or of the histological type, each gynecological tumor showed a different degree of sensitivity. The results suggested that it is necessary to choose an oral antitumor agent according to the degree of sensitivity in oral adjuvant chemotherapy. In an experimental study with an implanted tumor, 14 of 20 mice (70.0%) developed a recurrent tumor in the control group. Compared to this, the recurrence rates for tumors in mice treated with CPM, CQ and UFT were 10.0% (p less than 0.001), 25.0% (p less than 0.01) and 30.0% (p less than 0.02), respectively. All these agents were considered to be effective in preventing tumors from recurring. In SRCA, the implanted tumor was sensitive to CPM and CQ, but not sensitive to UFT. These data suggested that SRCA is useful in predicting the effect of dose-dependent agents in oral adjuvant chemotherapy, although, with UFT, a time-dependent agent, it is not clear whether SRCA is appropriate for estimating the usefulness of its agent.     

Laparoscopic excision of ovarian neoplasms subsequently found to be malignant.

One hundred fifty-six members and candidate members of the Society of Gynecologic Oncologists responded to a survey concerning the "laparoscopic management of ovarian neoplasms subsequently found to be malignant." Twenty-nine responders (19%) reported a total of 42 cases of ovarian malignancy. The laparoscopic procedure was aborted or the cyst was aspirated in 38% of the cases, and partial or complete excisions were attempted in 33 and 29%, respectively. The characteristics of the masses were as follows: less than 8 cm 67%, cystic 62%, unilocular 48%, and unilateral 81%. All four "benign" characteristics were present in 31% of the cases found to be malignant, and three of four characteristics were present in 24%. Laparotomy was performed at the time of laparoscopy in 17% of cases, after laparoscopy in 71% with an average interval of 4.8 weeks, and not at all in 12%. Fifty-seven percent of the cases were invasive epithelial malignancies, whereas 29% were tumors of low malignant potential. At least 50% of the patients had stages II-IV. We conclude that attempted laparoscopic excisions of adnexal masses that are subsequently found to be malignant are not uncommon, and that the presence of so-called "benign" characteristics does not preclude malignancy. Attempts at partial or complete excision are common, as are delays in subsequent definitive surgery. The stage of disease is often advanced, and all histologic types of malignancy are encountered. We advocate careful evaluation of this practice, with development of strict guidelines to ensure optimal patient care.     

The diagnostic value of multiple tumor markers in malignant ovarian neoplasms

Objective:To study the diagnostic value of multiple tumor markers in malignant ovarian neoplasm.Methods:Sera obtained from 430 patients with ovarian masses (110 cases were malignant ovarian tumors,320 cases were benign ovarian tumors) before operation,and from 50 healthy women as control.Serologic examination of tumor markers included CA125,TSGF,SA,CEA,AFP,HCG and Fer.Results:The serum levels of CA125,TSGF,SA and Fer in patients with ovarian cancer were higher than those in patients with benign ovarian tumors (P<0.05),also in control group (P<0.05).In the diagnostic value of application for malignant ovarian neoplasm,CA125,TSGF and SA were better than the others.The sensitivity,specificity and accuracy in diagnosis of ovarian cancer were 86.4%,82.8%and 83.7% respectively for CA125 alone,78.2%,81.3%and 80.5% for TSGF alone,74.5%,81.9%and 80.0% for SA alone,whereas 95.5%,45.6%and 58.4% for multiple tumor markers combined in which 1 or more indices showed positive,93.6%,80.6%and 84.0% for that in which 2 or more indices showed positive,and 87.3%,90.3%and 89.5% for that in which 3 or more indices show positive.Conclusion:multiple tumor markers examination could improve the diagnosis of ovarian cancer,and examination of CA125,TSGF and SA combined is most ideal.