Search

We employed interstitial brachytherapy using a high dose rate Ir-192 remote afterloading unit in two breast cancer patients with locoregional recurrence. In the first case, skin metastasis was treated, with favorable control of the infield tumor but subsequent persistent sequelae and multiple outfield metastases. This experience caused us to be cautious when choosing brachytherapy for the second case, in whom a solitary metastasis to an axillary lymph node was successfully treated. Although this method is still investigational, it may play a critical role in the treatment of locoregional recurrence resistant to other treatment modalities.

Predictive markers and variables for response to anticancer therapy provide cancer patients with refinement of therapeutic options and a decreased likelihood of receiving an ineffective therapy. The best-established predictive marker for response to endocrine therapy for breast cancer is the status of estrogen receptors (ER) in the primary breast tumor. However, although patients with ER-positive tumors have a greater than 50% objective response rate to endocrine therapy, other patients can not obtain an objective response. Therefore additional markers, such as better molecular biologic markers, are needed. Our previous study using multivariate analysis revealed that the ER status of primary tumors and the dominant site of metastasis are independent predictors for response to first-line endocrine therapy and that a response to first-line endocrine therapy is only an independent predictor for response to second-line endocrine therapy. However, all these factors are already well-established predictive markers for response to endocrine therapy. Recently, a number of new hormonal agents, such as more selective aromatase inhibitors and specific antiestrogens, have been developed and introduced. However, several questions, such as the best sequences when using hormonal agents, remain to be elucidated. On the other hand, several molecular biologic markers predicting response to endocrine therapy, such as the expression of the HER family of tyrosine kinase receptors, pS2, Bcl-2, and vascular endothelial growth factor, have been reported. To elucidate the most effective use of endocrine therapy for recurrent breast cancer, classical and new predictive factors for response to endocrine therapy are reviewed, and the clinical implications of these factors are discussed.

BACKGROUND: Four different endocrine therapeutic agents have been used in Japan since 1996. However, a consensus regarding proper use of these agents has notyet been established. Therefore, a questionaire survey of Japanese breast cancer authorities on endocrine therapy and a multi-institute survey to investigate the efficacy of a single first-line endocrine therapy for recurrent breast cancerwere conducted. MATERIALS AND PATIENTS: A total of 279 questionaires were sent to the Councilors of the Japanese Breast Cancer Society. The clinico-pathological data of 77 breast cancer patients who underwent a single first-line endocrine therapy were collected from five institutes. RESULTS: The response rate to this questionaire survey was 67.4%. The resultsshow that many authorities consider that: 1) both ER and PgR in primary tumors should be measured, 2) patient age, the disease-free interval and postoperative adjuvant therapy do not provide enough information for the selection of endocrine therapies, 3) antiestrogen and LH-RH agonists should be used as first-line endocrine therapies, 4) combined endocrine therapies, such as an antiestrogen plus an LH-RH agonist, should be used, 5) the optimal sequence of use of endocrine therapeutic agents is most controversial. The objective response rate to first-line endocrine therapies was 40.3% and the duration of response was over 15 months.The objective response rate to second-line endocrine therapies was 42.1%. A multiple regression analysis of predictive factors for the efficacy of first-line endocrine therapies indicated two factors, the disease-free interval and dominantsite of metastasis, to be significant. Conclusions: This questionaire survey suggests that clinical trials to investigate the optimal sequence of use of endocrine therapies and to clarify the usefulness of combined endocrine therapies should be conducted. Single first- or second-line endocrine therapies for recurrent breast cancer are effective and should be carried out by general clinicians.

We retrospectively analyzed the outcome for patients with locally invasive papillary carcinoma. The study group comprised 40 patients with locally invasive papillary thyroid carcinoma first diagnosed between 1981 and 1995. The enrolled patients were divided into two groups according to whether they underwent complete resection (n = 19) or not (n = 21). All patients were followed-up for a maximum of 206 months and a minimum of 33 months until 1998. There were no significant differences among these two groups with regard to age, sex, or tumor size. Recurrence of the disease was recognized in four patients in the complete resection group and ten in the incomplete group. There were no significant differences in the recurrence rate between both groups. Five patients from the incomplete resection group died of disease, and all patients from the complete resection group were alive. The percentage of surviving patients in the complete resection group was significantly higher than that in the incomplete group. The 15-year survival rates of the complete resection group and incomplete resection group were 100% and 74.2%, respectively. The 15-year survival rate of patients younger than 45 years in the incomplete resection group was 100%. The 15-year survival rate of the complete resection group was significantly higher than that of the incomplete group. In conclusion, complete resection without tumor residue should be performed for patients older than 45 years.

Vascular endothelial growth factor (VEGF)-A is known to play an important role in tumor angiogenesis. Three additional members of the VEGF family, VEGF-B, -C and -D, have recently been discovered. VEGF-C and VEGF-D are ligands for VEGF receptor-3, which is expressed in the endothelium of lymphatic vessels. The expression of VEGF-C is known to be associated with the development of lymphatic vessels. Therefore, it is conceivable that VEGF-C and VEGF-D might play a role in the development of lymphatic vessels in solid tumors. To obtain some clue as to this role, we developed a semi-quantitative reverse transcription-polymerase chain reaction method to investigate the mRNA expression levels of each VEGF family member in breast cancer. All the VEGF family members were expressed at different levels in seven human breast cancer cell lines explored. Although VEGF-A and VEGF-B expressions were detected in both node-positive and node-negative breast tumors, VEGF-C expression was detected only in node-positive tumors. VEGF-D expression was detected only in an inflammatory breast cancer and a tumor which developed an inflammatory skin metastasis. These findings suggest a possible relationship between the expression level of VEGF-C and/or VEGF-D and the development of lymphatic tumor spread.

Cancer cachexia is a common paraneoplastic syndrome in patients with advanced malignancies. However, the mechanisms of the development of cancer cachexia remain to be elucidated. Interleukin (IL)-6 is known to be involved in the development of cancer cachexia. The KPL-4 human breast cancer cell line, which was recently established in our laboratory, secretes IL-6 into the culture medium. Oral administration of doxifluridine (5'-DFUR, 60 mg/kg or 120 mg/kg) significantly inhibited the growth of KPL-4 tumors, reduced the tissue levels of IL-6, and alleviated body weight loss. Serum IL-6 levels were also lowered by 5'-DFUR in nude mice bearing KPL-4 tumors. Additionally, it is suggested that tumor necrosis factor (TNF)-alpha is involved in the cachexia induced by KPL-4 tumors. We suggest that 5'-DFUR suppresses cancer cachexia by lowering IL-6 levels in the tumor tissues and serum.

We report a case of adenoid cystic carcinoma (ACC) of the breast with a prominent basaloid feature. The patient was a 62-year-old Japanese woman with a right breast mass, measuring 1.5 cm in diameter. Histologically, the tumor was composed of basal cell-like tumor cells, and it was originally diagnosed as invasive ductal carcinoma. The presence of PAS-positive basement membrane material around the tumor cell nests may be a diagnostic clue to ACC. The prognosis of ACC of the breast is considered to be favorable. However, basaloid type ACC may represent a poor prognosis, since our case revealed an aggressive behavior in spite of its small size.

The critical mechanisms responsible for antiestrogen resistance have not yet been elucidated. We previously established a breast cancer cell line, KPL-1, derived from a patient with recurrent disease which appeared under tamoxifenadministration. In a previous study, we suggested that this cell line is estrogen receptor (ER)-positive but tamoxifen-resistant. In the present study, the effects of a pure antiestrogen, ICI 182,780, on this cell line were investigated. Although tamoxifen inhibited neither cell growth nor estradiol-stimulated transcriptional activity in vitro, ICI 182,780, significantly inhibited both of them. Tamoxifen and ICI 182,780 were then administered to female nude mice bearing KPL-1 tumors. Tamoxifen had no effect on tumor growth, but ICI 182,780 unexpectedly stimulated it (p = 0.022). Estradiol tended to inhibit tumor growth (p = 0.198). Immunohistochemical analysis revealed that ICI 182,780 significantly increased the Ki6-labeling index (p<0.001) but estradiol decreased it (p = 0.035). To explore the possible mechanisms of these phenotypes, the mRNA levels of ER-alpha,ER-beta, transforming growth factor-beta1, fibroblast growth factor (FGF)-1 and FGF-4 in KPL-1 cells were compared with those in other ER-positive human breast cancer cell lines by reverse-transcription polymerase chain reaction. FGF-1 was overexpressed only in KPL-1 cells. This cell line is the first breast cancer cell line to be growth-stimulated by ICI 182,780 in vivo. Paracrine interaction between tumor cells and stromal cells mediated by growth factors, such as FGF-1, might be a key factor to explain the unique hormone responsiveness of KPL-1 cells.

To evaluate the effects of dietary fats on breast cancer growth and metastasis, KPL-1 human breast carcinoma cells which have a propensity for axillary lymph node metastasis when inoculated into the thoracic mammary fat pad of female nude mice were examined. The mice were fed one of three semipurified diets containing 9.5% eicosapentaenoic acid plus 0.5% linoleic acid (EPA diet), 10% linoleic acid (LA diet), or 9.5% palmitic acid plus 0.5% linoleic acid (PA diet), or commercial laboratory chow containing 8.5% fat of which 4.1% was LA, 1.1% was PA, 0.06% was EPA, and 3.24% was other (Standard diet) starting 19 days before tumor cell inoculation and continuing until the end of the experiment (43 days after tumor cell inoculation). The tumor growth was faster and at a higher incidence in the mice fed the LA diet, and much slower and at a lower incidence in the EPA diet group compared with the mice fed the PA or Standard diet; the two separate experiment demonstrated identical results. The differences in tumor weight between the LA and PA groups and between the PA and EPA groups were significant (P < 0.05, respectively) at the termination of the experiment; the differences were due to different tumor cell proliferation rates. In an in vitro MTT assay, fatty acids showed direct stimulatory or inhibitory effects on the KPL-1 cells. Lymph node metastasis was seen in the LA and Standard diet groups, whereas it was not seen in the PA or EPA groups. The body weights were significantly lighter in the LA and EPA groups compared with the PA and Standard diet groups (P < 0.05, respectively). The results indicate that the EPA diet produced a reduction in tumor cell growth and metastasis whereas the LA diet had an enhancing effect on these parameters; dietary fatty acids may thus have a direct role in the growth and metastasis of human breast carcinoma independent of their systemic effects.

Antitumor and antimetastatic activity of the angiogenesis inhibitor O-(chloroacetyl-carbamoyl) fumagillol (TNP-470), a semisynthetic analogue of fumagillin, was evaluated in breast cancer cell lines. In an in vitro MTT assay, after 72 hrs continuous exposure to TNP-470, growth inhibition was observed in all seven cell lines of murine (JYG-A, JYG-B, DD-762, and BALB/c-MC) or human (KPL-1, MDA-MB-231, and MKL-F) origin, in which the 50% inhibitory concentrations (IC50) at 72 hrs treatment were 4.6, 4.4, 4.6, 10.1, 35.0, 25.3, and 33.4 micrograms/ml, respectively. In an in vivo assay using JYG-A, JYG-B, KPL-1, and MDA-MB-231 cells by orthotopic (right thoracic mammary fat pad) transplantation in female nude mice, TNP-470 at 30 or 50 mg/kg body weight was injected s.c. every other day from the day of tumor cell inoculation until the end of the experiment. The inhibitory effect on primary tumor growth was obtained in all four cell lines in a dose-dependent manner. In the 50 mg/kg TNP-470-treated group, the reductions in tumor weight of the JYG-A, JYG-B, KPL-1, and MDA-MB-231 cells with respect to the controls were 50%, 30%, 4%, and 49%, respectively. Metastasis was seen in the JYG-A, JYG-B, and KPL-1 cells. The numbers of mice bearing pulmonary metastases of JYG-A and JYG-B cells and regional axillary lymph node metastases of KPL-1 cells were reduced, and TNP-470 at the 50 mg/kg dose to KPL-1 cells significantly reduced lymph node metastases compared with the control. Although the weight gain was retarded in the TNP-470-treated mice, weight loss was not seen. TNP-470 was highly effective in the treatment of breast cancer cells. These results suggest that the clinical use of TNP-470 may be a promising treatment for breast cancer patients.

We recently established a metastasis model in nude mice using the MKL-4 cell line, a contransfectant of the MCF-7 human breast cancer cell line with fgf-4 and lacZ in which micrometastases in several organs can be quantitatively observed. First, to develop a new postsurgical metastasis model, we investigated the timing of occurrence of micrometastasis and the influence of tumor removal on the progression of micrometastasis in this model. Micrometastases into lymph nodes and lungs were detected 3 weeks after the cell injections. Tumor removal 3 weeks after the injections significantly enhanced the progression of micrometastasis into lymph nodes and bone. Second, to study the effect of a mixed compound, UFT (a molar ratio of uracil:tegafur of 4:1), which has been widely used in the postsurgical adjuvant setting in Japan, 15 or 20 mg/kg UFT were administered p.o. for 4 weeks to tumor-bearing mice or to mice in which transplanted tumors were resected 3 weeks after the injections. Either dose of UFT significantly inhibited the tumor growth as well as the progression of micrometastasis into lymph nodes, lungs, liver, and brain. In addition, enhanced progression of micrometastasis in all explored organs by the tumor removal was significantly inhibited by the administration of either dose of UFT. In conclusion, this new postsurgical metastasis model may be useful for evaluating the efficacy of agents used in the postoperative adjuvant setting. UFT may be an effective drug for inhibiting the progression of micrometastasis after surgery.

Invasion-inhibiting factor 2 (IIF-2) and its albumin conjugate have been reported to inhibit spontaneous metastasis of highly metastatic cancer cells with no effect on primary tumor growth. To confirm the inhibitory effects of the IIF-2 conjugate on tumor invasion and spontaneous metastasis, we administered the conjugate intra-peritoneally (i.p.) to female nude mice bearing transplanted tumors with MKL-4 cells, which are MCF-7 human breast cancer cells cotransfected with fibroblast growth factor 4 and lacZ. Neither 10 nor 20 mg/kg doses of the conjugate caused any inhibition of primary tumor growth, but 20 mg/kg significantly inhibited tumor invasion and spontaneous metastasis. Tumor invasion was measured by a novel computer-assisted image analysis. Spontaneous microscopic metastases into lymph nodes and distant organs were measured by whole organ staining for beta-galactosidase activity and observed with a dissecting microscope. The dose of 10 mg/kg significantly inhibited tumor invasion but not metastasis. Interestingly, the number of factor VIII-positive microvessels in the tumors was not reduced by treatment at either dose level. These findings suggest that the anti-invasive effect of the IIF-2 conjugate may reduce both lymphatic and hematogenous metastases in this MKL-4 metastasis model without affecting angiogenesis.

Serum carcinoembryonic antigens (CEA), CA 15-3, and tissue polypeptide antigens (TPA) have been used in monitoring the clinical course of patients with breast cancer. However, recent reports have suggested that the serial levels of these markers during therapy do not always correlate with the response to therapy. To clarify the usefulness of the serial combination assay of these markers in monitoring the clinical course of patients during therapy, we investigated the relationship between the initial changes and the kinetic patterns of the markers after therapy and the objective responses. When an increase or decrease of over 20% in these markers is taken to be significant, then the initial changes in all three markers significantly correlated with the therapeutic responses (P < 0.01). Five distinct kinetic patterns in the marker levels were observed. A paradoxical kinetic pattern of CEA and CA 15-3 levels--that is, an "initial surge and subsequent drop"--was seen in one-third of the responders. The TPA levels tended to exhibit a "steady decline" pattern in those responders. The sensitivity and specificity of the kinetic patterns to predict the clinical courses were significantly higher than those obtained from the analysis of initial changes. These findings thus suggest that adequate knowledge of the unique kinetics of each marker may help to make a more accurate prediction of the therapeutic responses.

Progression of breast cancer from an estrogen-dependent, slowly growing tumor amenable to tamoxifen treatment to an aggressive, metastatic, estrogen-independent phenotype has been mimicked by the transfection of MCF-7 breast carcinoma cells with fibroblast growth factors 1 or 4. FGF-transfected cells are aggressively tumorigenic in ovariectomized or tamoxifen-treated nude mice, conditions under which the parental cells would not produce tumors. When detection of metastasis was enhanced by lacZ transfection, the FGF-transfected MCF-7 cells were reliably metastatic to lymph nodes and frequently metastatic to lungs, in further contrast to parental cells. An antiangiogenic drug, AGM-1470, given to mice bearing tumors produced by FGF-transfected MCF-7 cells, produced a decrease in tumor size. The decreased tumor size was not as marked as that produced by treatment with pentosan polysulfate, an agent which would abrogate all autocrine or paracrine effects of the transfected FGF. Thus, increased angiogenesis may be a component of the phenotypic change produced by the FGF transfection, but other autocrine or paracrine effects may also be important. Since a clonal FGF-4 and lacZ doubly-transfected cell line, MKL-4, progressively lost expression of the transfected lacZ gene in individual cells, we performed successive rounds of fluorescence-activated cell sorting to select high-expressing cells. High-expressing cell populations thus obtained rapidly lost expression of beta-gal activity in continued culture. High beta-gal expressing clonal cell lines of MKL-4 cells established by either one or two rounds of low-density cloning also lost lacZ expression with continued culture. Southern analysis of DNA from lacZ transfected cell lines showed the transfected sequences to be present and grossly intact in both high and low expressing populations. However, Northern analysis revealed that high-expressing populations of MKL-4 cells contained the most lacZ mRNA, implying that in the unstable MKL-4 cell line, individual cells are down-regulating mRNA levels of lacZ. Stable lacZ expression has been obtained in other FGF-transfected and parental MCF-7 cell lines using the same expression vector. Thus, the MKL-4 cell line is down-regulating mRNA encoding the transfected gene through a mechanism not dependent on the CMV promotor utilized in the expression vector. This evidence suggests that lacZ expression is not a benign modification in certain cells.

From January 1987 to December 1993, Stage I (T1N0M0) breast cancers were treated by quadrantectomy with axillary lymph node dissection plus radiation therapy (QUART) in 57 cases and by modified radical mastectomy (MRM) in 57 cases. The results for these two treatment groups were compared retrospectively. Booster irradiation of 10 Gy to the tumor bed was given to 1 of 5 cases with a positive resection margin. The remaining 56 cases received 50 Gy lineac X-ray to the ipsilateral breast. Systemic adjuvant therapy, tamoxifen and /or 5-FU derivatives p.o., were given to the majority of cases in both groups. There were no significant differences between the two groups with regard to the patient background; ie, age, tumor size, pathological node status, histology and estrogen receptor status. During the follow-up period of 12 to 89 months (median 55 months) in the QUART group, no local recurrence was observed and only 1 case of bone metastasis occurred. In the MRM group, recurrence occurred in 4 cases during the follow-up period of 12 to 95 months (median 52 months). These consisted of 2 cases of regional lymph node recurrence and 2 cases of lung metastases. The differences in the disease-free survival rate and the overall survival rate between the two groups were not significant. In the QUART group, mild radiation pneumonitis and mild telangiectasia in the breast skin were observed in 2 cases(3.5%) and 1 case, respectively. Mild arm edema was observed in 4 cases (7%) in each group. These results demonstrated that QUART was as effective as MRM in treating T1M0M0 breast carcinoma in Japanese women.

The quality of life (QOL) in 55 early-stage breast cancer patients after surgery was prospectively assessed using a newly developed Japanese QOL questionnaire: The QOL Questionnaire for Cancer Patients Treated with Anticancer Drugs (QOL-ACD). The impacts of breast conserving treatment (BCT) (22 cases) and modified radical mastectomy (MRM) (33 cases) on the QOL in those subjects were compared. The overallQOL scores were evaluated during four periods (before surgery, 0-2, 3-12, and 13-24 months after surgery). The mean scores of the four categories of the QOL-ACD (activity, physical condition, psychological condition, and social relationships) were also compared. The results demonstrated that a significant improvement was observed in the overall QOL scores among the three periods after surgery (0-2, 3-12, and 13-24 months) only in the BCT group (p<0.05). There were no significant differences between the two groups in the overall QOL scores during any of the three periods after surgery, and the mean score of the 'psychological condition' during 0-2 months period in the BCT group was significantly lower than that in the MRM group (p<0.05). These results suggest that BCT does not always improve the patients' QOL more than MRM does, and that the patients receiving BCT require more psychological support than those receiving MRM during the early postoperative period.

Systemic therapy, pain relief and the quality of life (QOL) of breast cancer patients with bone metastasis were described. QOL was measured using a "QOL questionnaire" developed by the Ministry of Welfare in Japan. It was proved objectively that QOL scores in the cases with bone metastasis were significantly low in terms of activity, physical psychological conditions. Chemoendocrine therapy, endocrine therapy and outpatient therapy showed a high QOL score. The cases with bone pain showed a low QOL scores. In the 45 cases whose first metastatic site was bone only, there were no differences between endocrine therapy and chemoendocrine therapy in the rate and period of response or the total QOL score. MPA showed a higher response rate and a higher pain relief rate than TAM. In the cases with bone metastasis but without severe visceral metastasis, MPA monotherapy showed an excellent response when the tumor was ER or PgR positive, or when there was a long disease-free interval of more than three years, or if there was no previous therapy. MPA monotherapy is a suitable firstline therapy in such cases. Radiation therapy was more effective for bone pain (response rate 96.3%), and it was also effective in cases in which systemic therapy was not.

Prospective studies have suggested that postoperative adjuvant chemotherapy for breast cancer resulted in a lower recurrence rate and a better survival rate. However, there have been little experimental data on the chemotherapeutic agents for the suppression of progression of micrometastasis from breast cancer focus. We recently established a spontaneous metastasis model of MKL-4 human breast cancer cells transplanted into nude mice. In the present paper, both antiproliferative and antimetastatic effects of UFT were studied with this metastasis model. Oral administration of UFT (20 mg/kg daily) given 4 weeks inhibited significantly the growth of MKL-4 tumors in mice. Histological examination revealed a massive necrosis in the treated tumors. UFT also seemed to decrease the frequency of lymph node and distant metastases. In particular, no metastasis was observed in tumor-bearing mice in which the volume of tumors after the UFT-treatment was smaller than that before the treatment. It is premature to conclude that UFT has an antimetastatic effect because the volumes of the tumors of the control group and the treatment groups differed, but at least UFT may have an antimetastatic effect.

It has been suggested that the timing of surgery during periods of unopposed estrogen circulation, when high blood levels of estrogen and low blood levels of progesterone exist, has a deleterious effect on the survival of premenopausal patients with breast cancer. We studied this controversial issue by examining the serum estradiol and progesterone levels of 38 premenopausal patients with primary breast cancer, and by analyzing data on 100 premenopausal patients treated for primary breast cancer. The survival of 31 patients who had undergone initial surgery between days 3 and 12 after their last menstrual period (group E) was compared with that of 69 patients who had undergone surgery between days 0 and 2 or from 13 days after their last menstrual period (group P). The overall survival of group E was significantly worse than that of group P (P = 0.049). This difference was especially notable in patients with node-positive tumors or tumors larger than 2 cm in size; however there was no significant difference in disease-free survival between the two groups. On a multivariate analysis, nodal status was the only significant prognostic factor for both overall and disease-free survival. Thus, these findings suggest that unopposed estrogen circulation may be detrimental to the overall survival of premenopausal women with breast cancer.

The objective of this study was to investigate the factors associated with the quality of life (QOL) in breast cancer patients after surgery. The QOL in 83 primary breast cancer patients after surgery was prospectively assessed using a newly developed Japanese QOL questionnaire: The QOL Questionnaire for Cancer Patients Treated with Anticancer Drugs (QOL-ACD). The demographic and medical factors relating to the QOL score in the subjects were investigated by multiple regression analyses. Factors related to the four categories of the QOL-ACE (activity, physical condition, psychological condition, and social relationship) were also analyzed. The results revealed that only hospitalization had a strong negative relation to the overall QOL score. With regard to the four categories of the QOL-ACE, hospitalization had strong negative relations to the three categories (activity, physical condition and social relationship) and had weak negative relation to the psychological condition. Other demographic and medical factors such as types of surgery or adjuvant therapy had no significant association with the QOL score, except for the performance status which had positive, but not significant, association with the activity. In conclusion, this study yielded useful information concerning the management of the breast cancer patients after surgery. We had better be more concerned with the hospitalized period than other demographic or medical factors such as types of therapy to improve the QOL.

Accumulated knwoledge regarding the important roles played by angiogenesis in solid tumor growth and metastasis has prompted the development of angiogenesis inhibitors for clinical use in cancer therapy. Among these inhibitors, O-(chloroacetyl-carbamoyl) fumagillol (TNP-470) has been reported to have both antitumor and antimetastatic activities in rodent and human tumor models. We recently established a new metastasis model of MKL-4 human breast cancer cells in female nude mice. This cell line is a co-transfectant of the MCF-7 cell line with genes of a potent angiogenic factor, fibroblast growth factor 4, and a genetic marker, bacterial lac ¤ In this paper, we describe the inhibitory effects of TNP-470 on tumor angiogenesis, growth and metastasis in this MKL-4 metastasis model. Subcutaneous injection of 10 or 50 mg/kg of TNP-470 every other day for two weeks obviously inhibited the tumor growth and metastasis of MKL-4 cells in female nude mice. The treatment of TNP-470 at both doses significantly reduced the tumor volumes, respectively, to 28% and 14% of that in the control group(p<0.05 in each comparison). The positive rate of metastasis into the axillary lymph nodes was 100% in the control group, whereas it was only 33% in both of the treatment groups. Distant metastasis into the inguinal lymph nodes, lungs, kidneys and liver also tended to decrease in both of the treatment groups. Immunohistochemical analysis using anti-factor VIII antibody revealed that the number of microvessels in both of the treatment groups was significantly less than that in the control group(p<0.01 in each comparison). To the best of our knowledge, this is the first report describing simultaneous demonstration of the antiangiogenic, antitumor and antimetastatic effects of TNP-470 on human breast cancer cells in nude mice.

Using a questionnaire survey, we analyzed the relationship between the frequency of breast self-examination (BSE) and the clinical stage and course of breast cancer in Japanese patients. BSE had been performed monthly by only 5.4% of the patients (M group), occasionally by 35.4% (O group), and not at all by 59.2% (N group). There was a positive relationship between more frequent BSE and an earlier clinical stage, the percentages of Tis/stage 0 and I for the M, O, and N groups being 83%, 44%, and 36%, respectively (P < 0.05). The mean maximum tumor diameters for the three groups were 1.7 cm, 2.5 cm, and 3.0 cm, respectively. The tumor size in the M and O groups was significantly smaller than that in the N group at P < 0.01 and P < 0.05, respectively. The percentages of patients in the M, O, and N groups who underwent breast-conserving therapy were 42%, 11%, and 19%, respectively, with patients who had performed monthly, BSE more frequently undergoing breast-conserving therapy (P < 0.05). At a median follow-up time of 34 months, 0%, 3.8%, and 7.6% of the patients from the M, O, and N groups, respectively, had died of breast cancer, the overall survival curve of the M group being significantly better than that of the N group (P < 0.01). This retrospective study suggests the positive correlation of BSE frequency with earlier detection, and a more favorable clinical course in Japanese breast cancer patients.

The characteristics and clinical efficacies of new hormonal drugs and new treatment methods for breast cancer using tamoxifen (TAM) were described. The new anti-estrogens, such as trioxifen, toremifene and droloxifen, have equal or better clinical efficacy than TAM. They showed a high response rate in postmenopausal patients with no previous treatment or estrogen receptor positive tumors. They were occasionally effective for patients who relapsed after TAM treatment. Pure anti-estrogen ICI 164, 384, with fewer estrogenic agonistic properties than TAM, and anti-estrogen TAT-59, which has been developed in Japan, are under clinical investigation. Luteinizing hormone-releasing hormone agonists, such as leuprolide, buserelin, triptrelin and goserelin, are available for premenopausal patients, and showed a 37% response rate on average. The pure aromatase inhibitors, such as 4-hydroxyandrostenedione and CGS 16, 949 A, are available for postmenopausal patients, and showed 30% and 17% response rates on average, respectively. The new pure aromatase inhibitor ZD-1033 is also under clinical investigation. TAM or toremifene as well as the calcium antagonist verapamil were proved to overcome the multiple anticancer drug resistance caused by P-glycoprotein. Clinical studies for breast cancer using high-dose TAM as a potential modulator of drug resistance have already been started in Europe and the United States. Chemoprevention of breast cancer using TAM, which started in the U.S., was described.

Successful antiestrogen treatment in patients with tamoxifen-responsive breast tumors is often followed by an outgrowth of tumors cells that are antiestrogen resistant, implying that estrogen-dependent tumors can become estrogen-independent. In an effect to mimic this progression, we have transfected fibroblast growth factor 4 into MCF-7 cells, a human breast carcinoma cell line that is estrogen-dependent for growth in nude mice. This transfection results in cell lines that form progressively growing, metastatic tumors when injected s.c. into untreated or tamoxifen-treated ovariectomized nude mice. In contrast to the parental cell line, growth of transfected cells in ovariectomized nude mice is stimulated by tamoxifen treatment and inhibited by estrogen treatment of the mice. Parental MCF-7 cells were transfected with an expression vector for beta-galactosidase, conferring the ability to convert the chromogenic substrate, 5-bromo-4-chloro-3-indoyl-beta-galactoside, to a blue color and allowing the detection of their presence within tumors developing after coinoculation with fibroblast growth factor 4-transfected cells. The fibroblast growth factor 4-transfected cells could support growth and metastasis of the beta-galactosidase-expressing parental cell line when both lines were coinjected into the same site in untreated or tamoxifen-treated, ovariectomized mice. These data suggest a possible role for fibroblast growth factors in the progression of breast tumors to an estrogen-independent, antiestrogen-resistant, metastatic phenotype. They also support a role for paracrine factors in mixed populations of tumor cells of differing states of malignant progression.

We recently established transfectants of MCF-7 human breast cancer cells with fibroblast growth factor 4 (fgf-4) that showed rapid growth and spontaneous metastasis in ovariectomized and tamoxifen-treated nude mice. To establish a spontaneous metastatic model of human breast cancer cells in nude mice with a sensitive marker for detection of micrometastasis, the transfection of fgf-4 was combined with transfection of the bacterial lacZ gene encoding beta-galactosidase. MKL-4 cells, a lacZ transfectant of an fgf-4-transfected cell line, showed the same level of fgf-4 expression as parental cells and expressed a high level of beta-galactosidase activity. When MKL-4 cells were injected s.c. into female nude mice, rapidly growing tumors developed. Whole organ staining for beta-galactosidase activity was able to detect even small numbers of metastatic tumor cells. Micrometastases in lymph nodes, lung, and brain were detected 3 weeks after the tumor cell injections, the first time point tested. Within 12 weeks, metastases were observed in lymph nodes, lung, brain, kidney, perirenal fatty tissues, liver, spleen, retroperitoneum, heart, and gallbladder. The frequency of metastasis and number of foci were correlated with the volume of the primary tumors. The distribution of metastatic sites was similar to that in breast cancer patients. MKL-4 cells may be a useful model for studying the malignant progression of hormone-dependent breast cancer, antimetastatic drugs, or early events in metastasis.

Both estrogen and progesterone receptors have been determined in 613 primary breast cancer patients treated by radical mastectomy. At the cut-off value of 5 fentomoles (fmol) cytosol protein/mg for both receptors, patients with estrogen or progesterone receptor-positive breast cancer showed significantly favorable disease-free, overall and post-recurrence survival curves to those of receptor-negative breast cancer patients. In the patient subgroups: premenopausal, stage III, more than four positive lymph node metastases, postoperative adjuvant tamoxifen therapy, a significantly favorable prognosis was recognized in either estrogen or progesterone receptor-positive patients. Both receptors are thought to be useful prognostic indicators for patients with advanced tumors or for those receiving postoperative adjuvant tamoxifen therapy. When the cut-off value was changed, the maximum significant difference in prognosis between receptor-positive and receptor-negative patients was observed at 5 fmol cytosol protein/mg for the estrogen receptor or 10 fmol/mg cytosol protein for the progesterone receptor. A more detailed examination should be made on the cut-off values of both receptors.

To encounter an apocrine carcinoma of the breast is very rare. We have experienced, however, 10 cases of an apocrine carcinoma (0.65%) among 1541 primary breast carcinoma cases. The mean age of the patients was 58.1 years, and the tumor size on palpation ranged from 1.1 to 6.0 cm (mean 3.5 cm). A lymph node metastasis was seen in 5 cases. While one patient determined as being in stage III b of the disease died of the cancer approximately 5 months after surgery, all of other patients are still alive. The prognosis of this type of disease appears to be better than that of a common invasive ductal carcinoma. Thirty-four cases of an apocrine carcinoma have been found in the Japanese literature, this number including our 10 cases. Discussed are the clinicopathological features and the histogenesis of this type of cancer.

Two rare cases of a tubular adenoma of the breast are reported, together with a clinicopathological review of the pertinent literature. Only 14 cases of this histological type have so far been reported in Japan. Both patients were single women aged 22 and 34 years, respectively. In case 1, a mass was found in the axilla, and in case 2, a breast cancer was simultaneously found in the contralateral breast. In each patient, a mass that measured 2 cm in diameter was oval, elastic and firm and smooth on the surface. In each instance the result of the biopsy performed led to a diagnosis of a fibroadenoma. Microscopically, the tumor was sharply demarcated from the adjacent breast tissue and consisted of homogeneous epithelial cells with small tubular structures and a minimal stromal component.

In order to investigate the mechanisms of the endocrine therapeutic agents and the applicability of combined endocrine therapies for breast cancers, we studied the effects of estrogen and the endocrine therapeutic agents on estrogen receptor (ER), progesterone receptor (PgR), and DNA synthesis in MCF-7 human breast cancer cells, which is known to be sensitive to estrogen. ER and PgR of MCF-7 cells were determined by whole cell uptake method. In brief, intact MCF-7 cells cultured in the multi-well plates were incubated with various concentrations of tritiated estradiol (E2) or promegestone (R5020) at 37 degrees C for 1 hour. The characteristics of the hormone binding were analyzed by Scatchard plots. MCF-7 cells had single class of ER (Kd: 2.1 +/- 0.2 X 10(-10) M, MBC: 9.0 +/- 1.5 X 10(3) sites/cell) and PgR (Kd: 7.2 +/- 1.1 X 10(-10) M, MBC: 3.1 +/- 0.5 X 10(4) sites/cell). When cultured in the presence of 10(-8) M or 10(-6) M of E2, tamoxifen (TAM), R5020 or medroxyprogesterone acetate (MPA) for 48 hrs, the numbers of binding sites of ER and PgR altered, but the affinities of either of them did not change. E2(10(-8) or 10(-6) M) increased about twice the number of PgR. Although treatment of 10(-8) M TAM, a non-steroidal antiestrogen, slightly increased the number of PgR, 10(-6) M TAM significantly decreased the number of PgR. Both of R5020 (10(-8) or 10(-6) M) and MPA (10(-8) or 10(-6) M), synthetic progestins, decreased the number of ER dose-dependently. On the other hand, E2 (10(-8) and 10(-6) M) and R5020 (10(-8) M) enhanced DNA synthesis, but 10(-6) M TAM or MPA inhibited DNA synthesis. The effects of single, sequential and coincidental treatment of these agents were compared. The sequential treatment of 10(-8) M E2 for 36 hrs and followed 10(-6) M MPA for 36 hrs ("10(-8) M E2----10(-6) M MPA") and "10(-6) M TAM----10(-6) M MPA" inhibited DNA synthesis of MCF-7 cells more efficiently than 10(-6) M TAM alone or 10(-6) M MPA alone for 72 hrs. However, "10(-6) M MPA----10(-6) M TAM" inhibited DNA synthesis less than "10(-6) M TAM----10(-6) M MPA".(ABSTRACT TRUNCATED AT 400 WORDS)

Two very rare cases of lipid-secreting carcinoma of the breast are reported, together with a review of the literature. Both patients were Japanese women aged 70 and 81 years, respectively. In each, a mass had been found in the breast, and radical mastectomies were performed based on a diagnosis of breast cancer at an early clinical stage. Both resected tumors were found to be solid and localized. In them, both estrogen and progesterone receptors were negative. Microscopically, hematoxylin-eosin staining showed the tumor cells to have foamy and vacuolated eosinophilic cytoplasm. Lipid staining showed the vesicles in the cytoplasm of the tumor cells to be colored a flaming red and consequently recognized as lipid droplets. An electron-microscopic study showed that the tumor cells contained various sizes of lipid droplets in the cytoplasm with no degenerative changes, and suggested that these lipid droplets were produced and stored in the tumor cells. It would appear, from the literature that lipid-secreting carcinomas have aggressive clinical courses, but neither vascular invasion nor lymph node metastasis was seen in either of our two cases.

In Gunma prefecture, mass screening for both breast and thyroid cancer at the same time was conducted between April 1980 and March 1988 with the cooperation of the Gunma Cancer Society. In this paper, the results of mass screening for breast cancer and further problems are reported and discussed. The total number of subjects during these eight years was 191, 603 on the first screening with physical examination alone. Of these, 18,208 (9.5%) examinees were required to attend the second examination. Breast cancer was detected in 159 cases. The detection rate was 0.08% among total subjects. Also thyroid cancer was detected in 244 (0.13%). The detection rate for breast cancer was approximately twice as high at the initial screening (0.11%) than at the subsequent periodic screening (0.06%). Patients with breast cancer detected by mass screening showed an earlier stage and a better prognosis compared with those of our ordinary outpatients. The 8-year disease-free survival rate in mass screening and in outpatient cases was 94.9% and 83.2%, respectively, but there was no significant difference between the two groups. Interval breast cancer was also found in 19 cases (Tis; 1, Stage I; 8 and Stage II; 10) and 5 patients had nodal metastases. This mass screening program is a useful means for early discovery and early treatment of breast cancer. Hereafter, we should intend to increase the number of initial examinees and women aged 60 and over. Also it is important to make and train physicians for such screening, and to educate women about the practice of breast self-examination.

Mass screening for detecting both thyroid and breast cancer in adult women was carried out in Gunma Prefecture, Japan, between 1980 and 1986. The total number of subjects during the seven years was 152,651. The first screening was by inspection and palpation using experienced surgeons. Thyroid abnormalities were found in 5,902 subjects (3.9%) and 3,804 (2.5%) were required to attend a second examination. Thyroid carcinoma was detected in a total of 216 subjects, a detection rate of 0.14%. The detection rate for thyroid cancer was twice as high at the initial screening (0.19%) than that at subsequent periodic screenings (0.09%). Histological classifications of the detected thyroid cancers were: papillary carcinoma, 189 cases (87.5%); follicular carcinoma, 26 (12%); medullary carcinoma, 1 (0.5%). Eighty-seven percent of the thyroid cancers were asymptomatic at the time of screening. The tumor size were 2 cm or less in 126 patients (58.3%). Thyroid cancer detected by mass screening was revealed at an earlier stage and showed a significantly better survival rate compared to that of our usual outpatients. As differentiated carcinoma has been found at a higher incidence by mass screening, the program is useful for the early detection and treatment of thyroid cancer.

Two very rare cases of a lipid-secreting carcinoma of the breast are reported, and a review of the pertinent literature. The patients were both women, aged 70 and 81, respectively. Both of their clinical stagings were T2aN1aM0, stage II., and radical mastectomies were performed. On examination of the resected specimens, the tumors did not differ from the general findings of breast cancer. Microscopically, H-E stain showed the tumor cells to have a foamy and vacuolated eosinophilic cytoplasm. Lipid stain and an electron microscopic study were performed and showed that the tumor cells contained large amounts of neutral lipid droplets in their cytoplasm.