H Tokita

Chiba Cancer Center, Tiba, Chiba, Japan

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Publications (21)28.49 Total impact

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    ABSTRACT: Background. The antitumor effect and toxicity of immunoconjugates were studied in nude mice bearing a human ovarian cancer cell line, OVA-1. Methods. We studied the tissue distribution of an anti-cytokeratin-8 monoclonal antibody (6D7) in OVA-1-bearing nude mice by giving 6D7 labelled with 125I. The immuno conjugate consisted of 6D7 and carboplatin (6D7-conjugate), coupled via carboxymethyl dextran, and this was intraperitoneally administered to OVA-1 bearing nude mice. The tumor volume and the body weight were measured for 5 weeks. Tissue platinum concentrations in the OVA-1 tumor, blood, liver, kidney, and spleen, were measured from 3 to 120 min after administration of the conjugate. The results were compared with those in nude mice treated with nonspecific mouse IgG coupled with carboplatin (IgG-conjugate) or carboplatin alone. Results. The coupling rate of the drug to 6D7 was approximately 80%, and was stable over several measurements at various times. In-vivo accumulation of 6D7 labelled with 125I in the OVA-1 tumors was significantly higher than that in mice that received nonspecific mouse- IgG-125I, with tumor/ blood radioactivity ratios of 14.0 and 1.28, respectively. The tumor growth rate in mice that were administered 6D7-conjugate was (at a maximum) 40% lower than the tumor growth rate in mice administered carboplatin. The body weight of the mice that received 6D7-conjugate did not decrease during the 5-week observation period, while the body weight of the mice that received carboplatin decreased by a maximum of 10%. In addition, upon administration of 6D7-conjugate, the platinum concentration in the tumor was maintained for a longer period than after the administration of carboplatin alone. Conclusions. The tumor growth suppression effect was significantly higher in the mice bearing the OVA-1 tumor that received 6D7-conjugate than in the animals that received carboplatin alone. This difference could be caused by differences in the platinum concentrations in the tumor between the two groups.
    International Journal of Clinical Oncology 07/1999; 4(4):236-240. DOI:10.1007/s101470050060 · 2.17 Impact Factor
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    ABSTRACT: A new cell line, Yumoto, derived from a squamous cell carcinoma of the uterine cervix, was established from serially transplanted tumor tissues in nude mice. Monolayer cultured cells were polygonal and formed pavement-like sheet. They showed a piling-up tendency and were devoid of contact inhibition. Electron micrographs demonstrated the presence of microvilli on the cell surface, abundant tonofilaments in the cytoplasm, and the connection with desmosomes. These electron micrographical characteristics of Yumoto cells were consistent with those of squamous cell origin. Yumoto cells were highly tumorigenic in BALB/c nude mice and produced a well-differentiated squamous cell carcinoma of keratinizing type which closely resembled to the original tumor tissues in nude mice. The presence of HPV DNA was examined using polymerase chain reaction and Southern blot analysis, but no known types of HPV DNA could be detected. Exons 2 through 11 of the p53 gene were analyzed by direct DNA sequencing, revealing a homozygous mutation at codon 281 in exon 8, GAC to CAC (Asp-->His). Furthermore, physical p53-gene deletion was demonstrated by dual-color fluorescence in situ hybridization. This cell line is useful for studying the carcinogenesis of cervical carcinoma and for investigating the biological characteristics of a HPV-negative and mutated p53 squamous cell carcinoma of the uterine cervix.
    Gynecologic Oncology 10/1998; 70(3):339-47. DOI:10.1006/gyno.1998.5072 · 3.69 Impact Factor
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    ABSTRACT: It has been well documented that caspase-1 (interleukin-1beta-converting enzyme, ICE) and its related cysteine proteinases such as caspase-3 (CPP32, apopain) and caspase-2 (ICH-1L) play important roles in apoptosis. In the present study, these genes were inserted into an inducible eukaryotic expression vector, pMSG, and transfected into NIH 3T3 mouse fibroblasts. The expression of caspases-1 and -3 was effectively induced by treatment with dexamethasone (Dex). The expression of caspase-2 was elevated in the transfected cells without treatment with Dex but was not further stimulated by Dex. High expression of these proteases alone induced neither apoptosis-like cell death nor any morphological change. However, the expression of caspase-1 but not of caspase-2 or -3 enhanced chemosensitivity toward cytotoxic anticancer drugs such as aclarubicin, epirubicin, adriamycin, nimustine and ifosfamide. It is thus concluded that caspase-1 mediates cytotoxic effects of these drugs.
    Anti-Cancer Drugs 02/1998; 9(1):82-7. DOI:10.1097/00001813-199801000-00010 · 1.89 Impact Factor
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    ABSTRACT: Interleukin-6 (IL-6) is a pleiotropic cytokine that is not only a mediator in major immunologic reactions but also a growth factor of keratinocytes. We studied the IL-6 secretion in vitro of 15 human cell lines derived from both squamous cell carcinoma (SCC) and adenocarcinoma of the uterine cervix. Four of the eight well differentiated SCC secreted a large amount (> 1500 pg/48 h/10(6) cells) of IL-6 in nude mice. In contrast, poorly differentiated SCC cell lines and all of the 7 adenocarcinoma cell lines secreted a small amount (< 500 pg/48 h/10(6) cells of IL-6). The expression of IL-6 mRNA of the cell lines correlated well with their IL-6 secretion potential. However, the expression of IL-6 receptor did not correlate with the IL-6 secretory potential. We also studied the IL-6 secretion of freshly isolated normal squamous epithelium and of dysplastic epithelium. In culture, two normal squamous epithelia secreted a large amount (> 2000 pg/48 h/10(6) cells), whereas 8 dysplasia epithelia secreted an extremely small amount (< 10 pg/48 h/10(6) cells). About one-third of patients with SCC had a raised serum IL-6 value. IL-6 production may help to differentiate between SCC and adenocarcinoma of the uterine cervix. IL-6 regulation seems to change in the course of SCC carcinogenesis.
    Archives of Gynecology and Obstetrics 02/1996; 258(1):25-33. DOI:10.1007/BF01370929 · 1.28 Impact Factor
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    ABSTRACT: Malignant fibrous histiocytoma is the most frequent soft tissue sarcoma. However, the pathogenesis still remains unclear, because there are very few human malignant fibrous histiocytoma cell lines available for precise cellular study. In this study, a human malignant fibrous histiocytoma cell line (MMF-1) was established from the pulmonary metastatic lesion of a 55-year-old man with malignant fibrous histiocytoma. Human cell line MMF-1 and its heterotransplanted tumor had almost the same characteristics as the original tumor morphologically and immunohistochemically. This cell line is expected to be a useful for studying the pathogenesis of malignant fibrous histiocytoma. The cloned cell lines (MMF-2 and MMF-3) also consisted of spindle-shaped, polygonal, and multinucleated giant cells, meaning that the fibroblast-like cells, histiocyte-like cells, and multinucleated giant cells seen in malignant fibrous histiocytoma were derived from a single tumor cell. Human cell line MMF-1 produced inflammatory cytokines, such as tumor necrosis factor-alpha, macrophage colony-stimulating factor, interleukin-8, and monocyte chemotactic and activating factor, that might be involved in the morphogenesis of malignant fibrous histiocytoma. Furthermore, the results of the analysis of human cell line MMF-1 suggested that malignant fibrous histiocytoma originated from a poorly differentiated fibroblast.
    Clinical Orthopaedics and Related Research 12/1995; 320(320):159-67. · 2.88 Impact Factor
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    ABSTRACT: Malignant fibrous histiocytoma is the most frequent soft tissue sarcoma. However, the pathogenesis still remains unclear, because there are very few human malignant fibrous histiocytoma cell lines available for precise cellular study. In this study, a human malignant fibrous histiocytoma cell line (MMF-1) was established from the pulmonary metastatic lesion of a 55-year-old man with malignant fibrous histiocytoma. Human cell line MMF-1 and its heterotransplanted tumor had almost the same characteristics as the original tumor morphologically and immunohistochemically. This cell line is expected to be a useful for studying the pathogenesis of malignant fibrous histiocytoma. The cloned cell lines (MMF-2 and MMF-3) also consisted of spindle-shaped, polygonal, and multinucleated giant cells, meaning that the fibroblast-like cells, histiocyte-like cells, and multinucleated giant cells seen in malignant fibrous histiocytoma were derived from a single tumor cell. Human cell line MMF-1 produced inflammatory cytokines, such as tumor necrosis factor-alpha, macrophage colony-stimulating factor, interleukin-8, and monocyte chemotactic and activating factor, that might be involved in the morphogenesis of malignant fibrous histiocytoma. Furthermore, the results of the analysis of human cell line MMF-1 suggested that malignant fibrous histiocytoma originated from a poorly differentiated fibroblast. (C) Lippincott-Raven Publishers.
    Clinical Orthopaedics and Related Research 10/1995; 320(320). DOI:10.1097/00003086-199511000-00027 · 2.88 Impact Factor
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    ABSTRACT: The experimental and clinical usefulness of a chemosensitivity test (Nuclear Damage Assay) was studied. Karyologic degenerative changes were observed as an indicator of drug sensitivity, in repeated arterial infusion chemotherapy (RAIC) using a reservoir for advanced hepatocellular carcinoma (HCC). In the experimental study, this sensitivity test was performed using five liver cell lines against 15 drugs. At the same time, the succinate dehydrogenase inhibition (SDI) test was also performed. Comparison of the results between these two tests gave a high consistency rate of 81%. Clinically, the karyologic sensitivity test was carried out in 135 patients with unresectable HCC. Drug sensitivity could be evaluated in as many as 89% of the total 135 patients. Of the patients, 43 received RAIC on an outpatient basis via a subcutaneously implanted reservoir. The objective response of RAIC on tumours of the 43 patients was evaluated as complete response, partial response, in 3 (9%) and 8 (23%) in 35 patients treated with positive drugs (positive group), and as 0 (0%) and 0 (0%) of 8 patients treated with negative drugs (negative group), respectively. As regards the prognosis, 1 year and 1.5 year survival rates were 70 and 45% in the positive group, and 42 and 0% in the negative group, respectively. As objective response in the positive group tended to be better than that in the negative group, and prognosis in the positive group was significantly better than that in the negative group, this sensitivity test appears to contribute to the improvement of therapeutic results if used to select drugs suitable for RAIC for advanced HCC.
    Journal of Gastroenterology and Hepatology 08/1995; 10(4):446-53. DOI:10.1111/j.1440-1746.1995.tb01598.x · 3.63 Impact Factor
  • N Isegawa · M Ohta · H Shirasawa · H Tokita · A Yamaura · B Simizu
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    ABSTRACT: The DNA genome of a canine oral papillomavirus (COPV) was completely sequenced and found to consist of 8607 base pairs, which were the longest of all known papillomaviruses (PVs). Its organization was similar to that of other PVs except that it lacked early gene 5 (E5) and possessed a unique long noncoding region (L-NCR) between the end of the early genes and the beginning of the late genes. COPV also possessed a short noncoding region (S-NCR) which contained a putative upper regulatory region (URR), which is commonly found in PVs. The L-NCR did not show any similarity to known PV DNAs nor other DNA sequences in the GenBank database. Nucleotide sequence analysis of COPV showed that it was closely related to human papillomavirus type 1 (HPV 1) and animal PVs associated with cutaneous lesions in rabbit, European elk, deer and cow as we reported previously.
    International Journal of Oncology 07/1995; 7(1):155-9. DOI:10.3892/ijo.7.1.155 · 3.03 Impact Factor
  • N Isegawa · K Nakano · M Ohta · H Shirasawa · H Tokita · B Simizu
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    ABSTRACT: Canine oral papillomavirus (COPV) DNA was isolated from two different sources. One of these DNAs was molecularly cloned and its physical map was determined. Hybridization analyses using subgenomic fragments of bovine papillomavirus type 1 (BPV-1) and human papillomavirus type 16 (HPV16) as probes revealed that the cloned COPV shared moderate homology within the E1 and L1 regions of BPV-1 and HPV16, whereas homology in other regions of BPV-1 and HPV16 was low. The putative L1 gene of COPV was sequenced and several conserved regions, including antigenic epitopes which are common in other known papillomaviruses, were analyzed.
    Gene 10/1994; 146(2):261-5. DOI:10.1016/0378-1119(94)90303-4 · 2.08 Impact Factor
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    ABSTRACT: At present, chemoterapy still achieves poor results in advanced pancreatic cancer and sometimes the patient's quality is impaired by useless chemotherapy. To improve this situation, we used the nuclear damage assay (NDA) as a chemosensitivity test for pancreatic cancer. We performed the NDA on percutaneous needle biopsy tissue obtained from 19 patients with inoperable advanced pancreatic cancer, and 16 of them were successfully assayed by the chemosensitivity test. We could get a positive response to the assay in 13 of the 16 patients. The 13 patients with a positive response were treated with the agent selected by the NDA, while 3 patients with no response were treated with 5FU. The outcome of chemotherapy was a complete response in one patient, no change in 5, and progressive disease in 7 of the patients showing tumor sensitivity in the assay. On the other hand, only progressive disease was found in the patients with no response in the assay. The survival rate of the former group was better than the latter (P<0.05), and the median survival time was 23.4 and 11.1 weeks, respectively. We also report in detail the patient with a complete response to VP-16, which is rarely effective against pancreatic cancer. These results suggest that NDA of percutaneous needle biopsy tissue might improve the efficacy of chemotherapy for advanced pancreatic cancer.
    Annals of Cancer Research and Therapy 01/1994; 3(2):103-108,70. DOI:10.4993/acrt1992.3.103
  • N Iijima · S Sekiya · H Tokita · T Oosaki · H Takamizawa
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    ABSTRACT: By means of a newly developed in vitro chemosensitivity test based on the morphological changes in the nucleus (nuclear damage assay) as previously described, we were able to screen currently available anticancer drugs within 24 hr with a 100% success rate. The nuclear damage assay was used to determine the chemosensitivity in 50 patients (66 assays) with ovarian cancer. The response rate for the 13 patients with measurable tumors, 8 of whom showed resistance to CAP (cyclophosphamide, adriamycin, and cisplatin) therapy, was 46 percent when they were given various combination chemotherapy protocols consisting of more than one active agent selected from group A and B agents by the nuclear damage assay. The newly developed in vitro chemosensitivity test proved to be useful when selecting a second line combination for patients with CAP-resistant ovarian cancer.
    Nippon Sanka Fujinka Gakkai zasshi 11/1991; 43(10):1323-8.
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    ABSTRACT: With a newly developed in vitro chemosensitivity test based on the morphological changes of nuclear damage (nuclear damage assay) described here, we were able to screen currently available anticancer drugs within 24 hr with a 100% success rate. In preclinical chemotherapy using four human ovarian epithelial tumor cell lines and their xenografts in nude mice, the in vitro/in vivo response (sensitive/sensitive and resistant/resistant) rate was 94%. The nuclear damage assay was used to determine the chemosensitivity in 49 patients (60 assays) with ovarian cancer. The response rate of the 13 patients with measurable tumors, 9 of whom showed resistance to CAP (cyclophosphamide, adriamycin, and cisplatin) therapy, was 46% when the patients were given various combination chemotherapy protocols consisting of more than one active agent selected from group A and B agents by the nuclear damage assay. The newly developed in vitro chemosensitivity test proved to be useful when selecting a second-line combination chemotherapy for patients with CAP-resistant ovarian cancer.
    Gynecologic Oncology 03/1991; 40(2):138-43. DOI:10.1016/0090-8258(91)90105-E · 3.69 Impact Factor
  • N Iijima · S Sekiya · H Tokita · H Takamizawa
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    ABSTRACT: By means of 3 different kinds of in vitro chemosensitivity testings--(1) a nuclear damage assay developed by us, (2) MTT assay, and (3) colony formation inhibition assay--we examined the sensitivity of 8 kinds of human ovarian cancer cell lines to various anticancer drugs. The sensitivity of in vivo xenografts of the cell lines in nude mice to anticancer drugs was also examined by inhibition of the tumor growth. The in vitro--in vivo correlation of sensitivity was studied in respect to both sensitivity and specificity rates. 1. Different active anticancer drugs were screened among the 3 in vitro chemosensitivity testings in the same human ovarian cancer cell line. 2. The in vitro--in vivo correlation of the nuclear damage assay (sensitivity 50%, specificity 94%) was the highest among the 3 testings. The nuclear damage assay which we developed therefore seemed to be the most useful assay method for clinical use.
    Nippon Sanka Fujinka Gakkai zasshi 03/1991; 43(2):191-6.
  • H Tokita · N Tanaka · T Ueno · S Fujimoto · S Sekiya · H Takamizawa
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    ABSTRACT: A simple in vitro sensitivity test of oncolytic drugs has been applied in 60 cases of human ovarian cancer. Tumor tissue blocks were minced with a razor blade, after which the cell clumps were poured into a tissue culture medium, containing an anticancer drug with a certain concentration, and incubated at 37 degrees C. Nine to 16 kinds of drugs were tested for each specimen. After incubation, the cell clumps were dispersed for providing smear specimens. Typical morphological changes appeared in the nucleus, characterized by karyorrhexis and karyopyknosis. The individual tumors displayed different sensitivities to the various drugs. The positive rate of alkylating agents ranged around 30%, and specimens examined after oncolytic treatment displayed low sensitivity.
    Gan no rinsho. Japan journal of cancer clinics 09/1988; 34(9):1101-5.
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    ABSTRACT: The patient was a 57-year-old woman with ovarian serous cystadenocarcinoma in FIGO clinical stage IV. Cancer antigen 125 (CA125), tissue polypeptide antigen (TPA) and carcinoembryonic antigen (CEA) were immunohistochemically demonstrated in tumor cells, and the variations of serum CA125 and TPA levels reflected the clinical course. The tumor tissue obtained at exploratory laparotomy was minced with scissors, and transplanted subcutaneously into female nude mice for in vivo maintenance. The tumor cells from 5th generation nude mice were dispersed in Eagle's minimal essential medium supplemented with 10% fetal calf serum, and incubated in Falcon tissue culture dishes at 37 degrees C in 5% CO2 in air for in vitro maintenance. The results were as follows: Histopathologically the tumor transplanted into nude mice showed a cystadenocarcinoma, which closely resembled the original human tumor. Immunohistochemically CA125, TPA and CEA were demonstrated in the tumor transplanted into nude mice as well as in the original human tumor. From the growth curve in nude mice, the doubling time was estimated to be about 3.5 days. Serum TPA levels in nude mice were increased in proportion to the tumor growth after transplantation, but serum levels CA125 and CEA were normal. The concentrations of CA125 and TPA were increased in the conditioned media compared with the control media, although the elevated values were decreased with subsequent passages. CEA concentrations in the conditioned media were unchanged.
    Archives of Gynecology and Obstetrics 02/1988; 243(2):69-81. DOI:10.1007/BF00932972 · 1.28 Impact Factor
  • H Tokita · N Tanaka · T Ueno · S Fujimoto · K Nakano
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    ABSTRACT: We have developed a simple in vitro chemosensitivity test for breast cancer. Tumor tissues were chopped finely with razor blade. The cell clumps were pounced into tissue culture medium, which contained a specified level of concentration of anticancer drugs, and incubated at 37 degrees C for four to eight hours. Nine to 10 kinds of drugs were tested on each specimen. After incubation, the clumps were pumped down. The cells were smeared and stained with Giemsa for microscopic examination. The individual tumors showed different sensitivities towards various drugs, and the typical morphological changes observed in their nuclei; were karyorrhexis and karyopyknosis.
    Gan no rinsho. Japan journal of cancer clinics 12/1986; 32(14):1803-8.
  • H Takamizawa · S Sekiya · H Iwasawa · H Ishige · H Tokita · N Tanaka
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    ABSTRACT: A new in vitro chemosensitivity test was developed from comparative studies on the cytotoxicity of anticancer drugs against human tumor tissues xenografted into nude mice and their cultivated cells in vitro. Half a gram of the material was sufficient to examine the sensitivity of the tissues to 10 kinds of potential anticancer drugs and the results were obtained within 24 hours. The test was applied to all of 20 patients with advanced ovarian cancer. The predictive accuracy was 58% in 12 evaluable patients. This response rate was higher than those of conventional combination chemotherapy with or without cisplatin and adriamycin. Individual ovarian cancers showed different sensitivities to the drugs. These results indicate that heterogeneity of sensitivity to anticancer drugs exists among individual ovarian cancers and that our new type of in vitro chemosensitivity test is useful for selecting the most effective drugs for each individual type of ovarian cancer.
    Gan to kagaku ryoho. Cancer & chemotherapy 01/1986; 12(12):2293-7.
  • T Katoh · H Ishige · H Takamizawa · H Tokita · N Tanaka
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    ABSTRACT: The effects of various anticancer agents on human choriocarcinoma transplanted to nude mice, specifically, inhibition effects on tumor growth and survival rate were studied to establish an appropriate chemotherapy for refractory choriocarcinoma. The agents studied were methotrexate (MTX), actinomycin D (ACD), cyclophosphamide (CPM), vincristine (VCR), L-PAM (MPI), bleomycin (BLM), carboquone (CQ), cisplatin (CDDP), ACNU, MCNU, vinblastine (VLB), VP16-213, OK-432, Maruyama vaccine (SSM) and metronidazole (ME), and the following results were obtained: 1) The inhibition effects on tumor growth were obtained in the groups of VCR, VP16-213, CDDP, MPL, CPM and CQ; 2) The survival rate was 100% in the groups of MPL, BLM, and CQ. In the groups of ACD VLB, VP16-213 and VCR, all mice died. 3) MPL was found to be the most effective agent in terms of inhibitory effect and survival rate. In the future, combination chemotherapy including MPL and maintenance chemotherapy with MPL to refractory choriocarcinoma should be studied.
    Gan to kagaku ryoho. Cancer & chemotherapy 06/1982; 9(5):843-8.
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    ABSTRACT: Experimental chemotherapy has been studying with choriocarcinoma in nude mice transplanted from drug resistant patients. The following results: 1) the combined therapy with MTX and ACD was not always effective and in place of one of them, the combination of CPM was effective 2) addition of VCR was effective; were obtained. These results were applied to two patients in clinic. 1) The first patient failed to obtain complete remission by the course of ACD, MTX + ACD, and MTX + ACD + CPM (MAC) therapy. Then the combination therapy with ACD + CPM made her HCG titer regressed to negative immediately. She is free from disease now. 2) The HCG titer and pulmonary metastasis of the second patient showed resistance to MAC therapy, CHAMOMA therapy, and the combined therapy with MTX. The combination chemotherapy with ACD + CPM + VCR made her HCG titer become to regressed and the pulmonary metastasis reduce to 1/4 in size. The clinical application of the results from experimental chemotherapy was defined to be possible. The experimental model will contribute to establish the schedule of the rational chemotherapy for drug-resistant choriocarcinoma.
    Nippon Sanka Fujinka Gakkai zasshi 03/1982; 34(2):173-9.
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    ABSTRACT: Serial transplantation in nude mice of human choriocarcinoma showed the drug resistance in clinic was succeeded and experimental chemotherapy has been studied to improve the prognosis of the patients with choriocarcinoma. The experimental tumors were two lines. CC-I-JCK (JCK) and CC-HM-I- (HM). In this paper, single and/or combination chemotherapy with methotrexate (MTX), actinomycin D (ACD), cyclophosphamide (CMP), and vincristine (VCR) were discussed. 1) Effects of the single agent: In JCK, the suppressive effects against tumor growth under administration of CPM or VCR, and some responses in histological finding were obtained. While, in HM, few suppressive effects against tumor growth and some responses in histological findings were obtained. 2) Effects of the combined agents: In JCK, the suppressive effects against tumor growth were as follows; nontreatment group less than MTX + ACD less than MTX + CPM less than MTX + ACD + CPM less than ACD + CPM less than ACD + CPM + VCR. The complete disappearance of tumors were gained under administration of the combination therapy with ACD and CPM. While in in HM, the effects were as follows; nontreatment group less than MTX + ACD less than ACD + CPM less than MTX + ACD + CPM less than MTX + CPM less than MTX + CPM + VRC. But no complete disappearance of tumor was obtained. 3) The good relationship between the results of experimental chemotherapy and the response of clinical patients who provided the tissues could be seen. It has been defined that the experimental chemotherapy with the tumors transplanted to nude mice in useful models for the establishment of rational chemotherapy for drug resistant choriocarcinoma.
    Nippon Sanka Fujinka Gakkai zasshi 01/1982; 33(12):2055-64.