[Effects of Olaparib, a PARP-1 Inhibitor, on Triple Negative Breast Cancer Cells with a BRCA1 Mutation].

In 2018, olaparib, a PARP inhibitor, was approved for the treatment of BRCA1/2 gene-mutation positive and HER2-negative inoperable and recurrent breast cancer; BRCA1/2 gene testing was also listed as a companion diagnostics. Here, we identified microRNAs(miRNAs) expressed after treatment with olaparib, which differed in the presence or absence of BRCA1 mutations in triple negative breast cancer(TNBC), and examined the changes in miRNAs after exposure to the combination of the PARP-1 inhibitor and a chemotherapeutic agent. After exposure to the PARP-1 inhibitor, the expression of miR-141, miR-155, miR-205, and miR-223 decreased in MDA-MB-231, HCC1143, and BT549 cells and increased more than 10 times in MDA-MB-436 cells. Moreover, the expression of miR-141 in MDA-MB-436 cells treated with the PARP-1 inhibitor together with gemcitabine increased more than 10 times; additionally, the expression of miR-205 increased more in the context of combination therapy versus single exposure to olaparib. The miR-200 family(including miR-141)and miR- 205 are known to function as ZEB1/2 targets and to act as epithelial-to-mesenchymal transition(EMT)-suppressors. Overall, these results suggest that it may be possible to recover the sensitivity of TNBC cells to chemotherapy via the suppressing EMT through the use of a PARP-1 inhibitor in the context of BRCA1 mutation.

[Evaluation of the Benefits of Administering Hand Therapy to Patients with Chemotherapy-Induced Peripheral Neuropathy].

Taxanes, which are used to treat breast cancer, damage the microtubules of normal nerve cells, causing numbness of the fingers related to chemotherapy-induced peripheral neuropathy(CIPN); therefore, effective methods for reducing numbness are needed. In 2017, it was reported that physical stimuli related to massage improved finger blood flow volumes, contributing to the regeneration of damaged nerves. We developed a method of hand therapy for breast cancer patients complaining of numbness related to anticancer drug administration, and examined its effects on numbness. Hand therapy was performed by a single therapist who received lectures at the Sophia Phytotherapy College, which is accredited by the Japan Handcare Association. The fingertips to wrist, ankle, metacarpal bones, palm, and elbow were massaged using the bilateral arms/fingers for 15minutes. We investigated the influences of daily living status(Support Team Assessment Sched- ule-Japan: STAS-J), age, body mass index(BMI), severity/site of numbness, type of numbness, type of drug, duration of breast cancer, duration of numbness, and presence or absence of lymph node dissection, and evaluated the severity of numbness using a 10-cm Visual Analog Scale(VAS). The study included 51 breast cancer patients complaining of numbness of the fingers, with a mean age of 59 years. In patients with relatively mild numbness(STAS-J 1), the VAS scores before and after hand therapy were 4.7±1.8 and 1.9±1.3, respectively, showing a marked decrease. In STAS-J 2 patients, the values were 4.9 ±1.4 and 2.1±1.3, respectively, also showing a marked decrease. Thus, this hand therapy reduced numbness in mild- and moderate-status patients. Statistical comparisons were performed between the STAS-J 1/2(mild/moderate numbness)and STAS-J 3/4(severe numbness)groups. Although the severity of numbness was not correlated with age, BMI, type of drug, lymph node dissection, or duration of breast cancer, the proportion of patients with a B1-year history of numbness was significantly larger in the STAS-J 1/2 group. The most frequent site of numbness was from the proximal interphalangeal joints to the fingertips. Concerning the severity of numbness, many patients complained of severe numbness, as represented by that after sitting straight. These results suggest that this hand therapy is effective for reducing numbness in patients receiving taxanes and complaining of mild to moderate numbness.

Bacterial profile of infant feces associated with lactation infectious breasts.

BACKGROUND: Mastitis is a common complication in lactating women. However, the diversity of intestinal bacteria in infant exclusively fed infectious milk remains uncharacterized. Our colleagues recently established a method based on 16S and 23S rRNA-targeted reverse transcription-quantitative PCR (RT-qPCR) for detecting bacteria. MATERIALS AND METHODS: In the present study, the bacteria present in 14 samples of milk and infant feces were characterized using the RT-qPCR method, and concentrations of fecal organic acids were measured during the period of breast massage using HPLC. RESULTS: Streptococcus agalactiae and Str. parasanguinis were detected in milk from mastitis patients, whereas Str. salivarius and Str. thermophilus were the predominant bacteria in milk from engorged breasts. In feces of breastfed infants, Str. salivarius, Str. thermophiles, and Str. parasanguinis were isolated. Levels of lactate were high in fecal samples, whereas the pH of infant feces stabilized during breast massage. The bacterial diversity of milk from lactation infectious breasts was similar to that in feces of infant fed milk from lactation infectious breasts. Streptococcus species isolated from the feces of breastfed infants are related to oral cavity health. CONCLUSION: These results suggest that Streptococcus species, which are part of the healthy oral microflora, may play an important role in preserving the intestinal bacterial flora in infants fed infectious milk.

Protein expression profile and microRNA expression signature in estrogen receptor-positive and -negative breast cancers: report of two cases.

BACKGROUND: Identification of the proteins that are associated with estrogen receptor (ER) status is a first step in selecting drugs against hormone-dependent breast cancer. Recently, the proteins associated with ER status were reported using liquid chromatography and tandem mass spectrometry, and microRNA (miRNA) profiling of breast cancer subtype was demonstrated using real-time-PCR. METHODS: We present herein two cases with differential protein expression and miRNA profiling in ER-positive and -negative breast cancer. RESULTS: Proteins associated with fatty acid metabolism were uniquely detected in ER-positive breast cancer. The level of miR-181a expression in ER-positive cancer was higher than that in ER-negative cancer, while the expression of miR-27a, miR-107, and miR-195 was lower in ER-positive compared with ER-negative cancer. CONCLUSION: These cases suggest that fatty acid synthase (FAS) and FAS-related miRNAs are important in ER-positive breast cancer.

Evaluation of the dislocation and long-term sonographic detectability of a hydrogel-based breast biopsy site marker.

PURPOSE: To evaluate the usefulness of the HydroMARK, a hydrogel-based breast biopsy site marker for ultrasound localization of breast lesions, we investigated the tendency for dislocation and sonographic detectability of the marker placed in patients. MATERIALS AND METHODS: The marker was placed in lesions that were expected to become obscured after biopsy for a suspicious breast lesion or after neoadjuvant chemotherapy for breast cancer. The patients consented to return for a repeat ultrasound ± mammography examination, and the degree of displacement of the marker was measured as the marker-to-residual lesion distance. RESULTS: The marker was placed after stereotactic biopsy, ultrasound-guided biopsy, and before/during neoadjuvant chemotherapy, in 11, 22, and 7 lesions, respectively. Surgical resection was performed for 22 of the 40 lesions, while remaining 18 benign lesions were followed. The marker was sonographically detectable in 89.7% (35/39), 100% (35/35), and 100% (18/18) of the cases, respectively, at a median of 8 days, 13 weeks, and 11 months after the deployment. The degree of displacement was lower in the ultrasound-guided placement group than in the stereotactic placement group (median displacement: 0 vs. 4.3 mm; p = 0.001), it was also lower in the core-needle biopsy and neoadjuvent therapy cases than in the vacuum-assisted biopsy cases (p = 0.003). At a median interval of 2.5 months after deployment, the marker remained unchanged in location in all cases (n = 18, p = NS). CONCLUSIONS: The HydroMARK appears to be a safe and effective marker with the advantageous characteristics of a low tendency for dislocation with time and long-term sonographic detectability.

Evaluation of the sonographic visibility and sonographic appearance of the breast biopsy marker (UltraClip®) placed in phantoms and patients.

PURPOSE: To evaluate the usefulness of the UltraClip® dual trigger breast tissue marker (UltraClip) for sonographic localization, we investigate the sonographic visibility and sonographic appearance of the UltraClip placed in phantoms and patients. MATERIALS AND METHODS: Ten UltraClips were placed in the target lesions in the phantoms. After the ultrasound examination of the UltraClip, the ultrasound images were compared to the real appearance of the UltraClip obtained by cutting the phantoms. In the patient, the UltraClip markers were placed after biopsy of a suspicious breast lesion or before or during neoadjuvant chemotherapy. The patients consented to return 1-3 weeks after the procedure for ultrasound imaging of the UltraClip. RESULTS: The UltraClip placed in the phantom appeared as a hyperechoic structure with a mean maximum diameter of 5.5 mm, which was found to correspond to the metallic clip in 90% (9/10) of the cases, and as a hyperechoic tubular structure with a maximum diameter of 9.0 mm corresponded to the expanded polyvinyl alcohol polymer in the remaining 10% (1/10) of cases. On the other hand, the UltraClip was detected as a hyperechoic structure measuring 3.5 mm in size only in 9 of the 15 (60%) patients. The sonographic visibility of the UltraClip was not affected depending on whether the target lesion or post-biopsy scar was sonographically detectable or not [60% (6/10) vs. 60% (3/5)]. CONCLUSIONS: While sonographic localization by targeting the UltraClip may be useful in 60% of the patients, another localization technique will be needed in the remaining patients.

Eribulin upregulates miR-195 expression and downregulates Wnt3a expression in non-basal-like type of triple-negative breast cancer cell MDA-MB-231.

Triple-negative breast cancer (TNBC), which does not show hormone sensitivity, is a poor prognosis disease without an established targeted treatment, so that establishing a therapeutic target for each subtype is desired. In addition, microRNA (miRNA), a non-cording RNA 19-25 nucleotide-longs in length, is known to be involved in regulating gene expression. We examined miRNA expression after exposure to eribulin, MDA-MB-231 cells, non-basal-like type of TNBC cell lines, and HCC1143 cells, basal-like type of TNBC cell lines. The activity of caspase-3 significantly increased compared to the control in MDA-MB-231, whereas no significant difference was observed in HCC1143. The expression level of 20-miRNAs significantly increased compared to the control in MDA-MB-231 after exposure to eribulin. The expression level of 6-miRNAs also significantly increased compared to the control in HCC1143. In these 2 cell types, miR-125b-1 and miR-195 were commonly expressed. While the expression level of miR-125b-1 decreased in both cells, the expression level of miR-195 increased in MDA-MB-231 and decreased in HCC1143. The expression level of miR-195 targeting Wnt3a significantly decreased compared to the control in MDA-MB-231, whereas it significantly increased in HCC1143. These results showed that exposure to eribulin highly increased the expression of miR-195 while it decreased the expression of Wnt3a in non-basal-like type of TNBC. Some miRNAs are known to regulate other signaling pathways involved in human pathogenesis by regulating the Wnt signaling pathway, and miRNA can act as a tumor-suppressing gene; therefore, miR-195 may serve as a therapeutic target in non-basal-like type of TNBC.

A case of Guillain-Barré syndrome with meningeal irritation.

Here, we report a 5-year-old girl with Guillain-Barré syndrome who presented with a chief complaint of pain in the extremities, which was followed by neck stiffness. Bladder dysfunction was found, which required catheterization. Magnetic resonance imaging revealed marked enhancement of the nerve roots in the cauda equina on T1-weighted imaging after gadolinium injection, and nerve conduction studies led to a diagnosis of Guillain-Barré syndrome. Her symptoms improved after intravenous immunoglobulin therapy, but her neck stiffness remained 16 days after admission. Four weeks after admission, she could walk without support. As patients with signs of meningeal irritation may be diagnosed with other diseases, such as meningitis, it is important to recognize atypical cases of pediatric Guillain-Barré syndrome to achieve early diagnosis and treatment.

The ß-catenin signaling pathway induces aggressive potential in breast cancer by up-regulating the chemokine CCL5.

ß-Catenin signaling plays a pivotal role in the genesis of a variety of malignant tumors, but its role in breast cancer has not been fully elucidated. Here, we examined whether deregulation of ß-catenin signaling is related to the aggressive characteristics of certain types of breast cancers. Analysis of cytokine levels in MDA-MB-231 cells overexpressing a constitutively active form of ß-catenin (CAß-catenin) revealed a higher level of CCL5 expression. Cells transfected with CAß-catenin or stimulated with recombinant CCL5 exhibited increased cell invasion activity and spheroid formation in vitro. Furthermore, CAß-catenin-transfected MDA-MB-231 cells formed larger tumor masses that contained more Ki-67-positive cells and infiltrating lymphocytes than did the control cells. An inhibitor of CCR5 and a pan-CXCR neutralizing antibody dramatically reduced CAß-catenin-promoted activities. In addition to CCL5, 6-BIO, a chemical activator of ß-catenin, induced cell invasion and spheroid formation in MDA-MB-231 cells. Furthermore, high levels of nuclear ß-catenin accumulation were detected in breast cancer in patients with metastasis but not in those without metastasis. Nuclear ß-catenin localization is related to increased CCL5 production in breast cancer. These findings suggest that ß-catenin expression enhances tumor progression via chemokine production in breast cancers and that ß-catenin signaling is a critical regulator of the aggressive traits of breast cancers.

Decreased miR-206 expression in BRCA1 wild-type triple-negative breast cancer cells after concomitant treatment with gemcitabine and a Poly(ADP-ribose) polymerase-1 inhibitor.

No targeted-therapy has been established for triple-negative breast cancer accompanied by mutations in breast cancer susceptibility gene1 (BRCA1) mutation. In the present study, using BRCA1 wild-type cells (MDA-MB-231) and BRCA1-mutated cells (MDA-MB-436), we investigated miRNA expression and apoptosis on day 1 after addition of gemcitabine-alone and in combination with poly ADP-ribose polymerase-1 (PARP1) inhibitor. After drug treatment, there were significantly fewer apoptotic BRCA1 wild-type cells than BRCA1-mutated cells. Expression of miRNA-26a, -29b, -100, and -148a increased in BRCA1 wild-type cells exposed to gemcitabine-alone and in combination with the PARP1 inhibitor. The addition of PARP1 inhibitor reduced miR-206 expression in BRCA1 wild-type cells but increased it in BRCA1-mutated cells. It was suggested that miR-206 serves as a target molecule of PARP1 inhibitor combination therapy for BRCA1 wild-type triple-negative breast cancer cells.

New device for MRI-guided surgical excision: report of a case.

A 37-year-old female was indicated to have a non-mass lesion in her left breast on ultrasonography (US) and visited our outpatient clinic. Mammography showed no findings of masses or microcalcification. Dynamic magnetic resonance imaging (MRI) showed a segmental enhanced lesion consisting of nodular and ring enhancement. A US-assisted vacuumed needle biopsy was performed, and the histological findings revealed sclerosing adenosis and apocrine metaplasia. After 1 year of follow-up, the MRI findings suggested both a benign lesion and ductal carcinoma in situ, and surgical excision was performed. We used a new device to evaluate the surgical margin on MRI. The non-mass lesion was excised according to the device-guided margin under local anesthesia. The histological findings revealed the features of mastopathy. Following excision, MRI showed no residual non-mass lesions, and the shape of the patient's left breast was maintained.

Gene analysis and dynamics of tumor stem cells in human glioblastoma cells after radiation.

Glioblastoma is the most malignant central nervous system tumor. Patients with glioblastoma are treated with a combination of surgery, radiotherapy and chemotherapy; however, this effect is not satisfactory with regard to the prognosis. It is reported that the tumor stem cells affect recurrence, and radio- and chemotherapy resistance of the tumor, and that these cells play an important role in tumorigenesis and tumor progression. Using human glioblastoma cell lines (T98G and A172), irradiated (0, 30, 60 Gy) glioblastoma cells were prepared under the same conditions as clinical therapy. We analyzed cell proliferation rate, side population analysis by fluorescence-activated cell sorting and isolation of CD133⁺ cells, and performed genetic analysis (human stem cells) on these cells. We also investigated the difference in gene expression in the cells after radiation. The stem cell-related genes were highly expressed in the CD133⁺ cells compared with the CD133⁻ cells, suggesting that the cancer stem cells may be located in these CD133⁺ cells. In the T98G cell line, the cell proliferation rate of 30-Gy irradiated cells was higher than those of non-irradiated cells and 60-Gy irradiated cells. Stem cell-related genes were highly expressed in 30-Gy irradiated CD133⁺ T98G cells. In conclusion, we suggest that CD133⁺ cells may strongly affect tumor proliferation and the resistance against radiation therapy.

Analysis of p53 and miRNA expression after irradiation of glioblastoma cell lines.

Glioblastoma is a malignant brain tumor that is difficult to completely cure by surgical treatment alone. However, resistance to anticancer drugs and radiation may be acquired during treatment. For instance, miRNAs involved in regulating the expression of genes inducing apoptosis and other specific genes have been proposed for use, in order to induce the apoptosis of radioresistant cancer cells. A172 glioblastoma cells, expressing wild-type p53 were irradiated to a total dose of up to 60 Gy allowing us to analyze the activities of apoptosis-related proteins. The miR-34a expression levels in cells after irradiation at 30 and 60 Gy were 0.17- and 18.7-times the BCL2 and caspase-9 expression levels, respectively. The high miR-34a expression level in the cells after irradiation at 60 Gy reduced the p53 expression level. This study suggests that apoptosis might be promoted by regulating the action of miRNAs, even in cells that have acquired radioresistance.

Health-related quality of life in patients with advanced colorectal cancer: results from a phase II study of S-1 combined with irinotecan (CPT-11).

BACKGROUND: We carried out this study to examine the health-related quality of life (HRQOL) of patients with advanced colorectal cancer treated with the oral fluoropyrimidine S-1 plus irinotecan (CPT-11). METHODS: HRQOL was assessed at baseline (pretreatment) and at 5-week intervals during treatment, using the European Organization for Research and Treatment of Cancer QLQ-C30 and QLQ-CR38 questionnaires. The HRQOL data for 12 preselected scales and 21 courses of treatment were then analyzed longitudinally. RESULTS: Thirty-seven patients completed the baseline and post-treatment HRQOL assessments. Statistically significant differences between the baseline and post-treatment HRQOL scores were observed for the global QOL, social function, and pain scales (all QLQ-C30), as well as the body image, future perspective, gastrointestinal tract symptoms, weight loss, and chemotherapy side effects scales (all QLQ-CR38); favorable post-treatment results were observed for all the scales except for body image and chemotherapy side effects, for which post-treatment deteriorations were observed. The changes in body image, future perspective, weight loss, and chemotherapy side effects were each greater than ten points and seemed clinically significant. CONCLUSION: Combined treatment with S-1 plus CPT-11 resulted in an acceptable deterioration in HRQOL functioning and symptoms, compared with baseline levels.

Health-related quality of life of colorectal cancer patients receiving oral UFT plus leucovorin compared with those with surgery alone.

BACKGROUND: Adjuvant chemotherapy of oral uracil/ftorafur (UFT) plus leucovorin (LV) has been accepted as the standard of care in the treatment of patients with stage II and III carcinoma of the colon. The objective of the study was to compare HRQOL reported by patients receiving oral UFT plus LV (UFT/LV group) versus no adjuvant treatment (control group) following surgery for colorectal cancer. METHODS: Ninety nine patients in the UFT/LV group and 83 in the control group participated. HRQOL was assessed with the European Organization for Research and Treatment of Cancer QLQ-C30 and HRQOL data measured longitudinally following surgery were compared between the groups. RESULTS: Eighty-eight percent (87 of 99) received all scheduled doses of UFT plus LV during the first three cycles, and 82 percent (81 of 99) did so for five cycles. The most common type of toxicity in the UFT/LV group was fatigue, which was generally mild. Six patients each had grade 3 diarrhea or anorexia. There were significant differences in the scores for role function, and specific limitations such as fatigue, nausea, and vomiting, dyspnoea, appetite loss, and financial difficulties, which deteriorated in the UFT/LV group. CONCLUSIONS: HRQOL in colorectal cancer patients with adjuvant chemotherapy with oral UFT plus LV deteriorated during this phase of treatment compared with those with surgery alone, despite the biased stage of tumor between the groups. Symptom management and social support would improve HRQOL in such a group of patients.

Evaluation of 5-fluorouracil related genes in breast cancer to predict the effect of adjuvant therapy with oral fluorouracil derivatives.

New anticancer drugs have been developed, and prediction of their effect is needed to perform tailor made chemotherapy. We investigated a selection of the predictive markers for oral adjuvant chemotherapy among 5-fluorouracil (5-FU) related genes. 5-FU related genes were examined by using a laser captured microdissection and real-time RT-PCR in 220 patients with invasive breast cancer. Sixty-six patients were treated with postoperative oral fluorouracil derivatives for 12 months or more, and we examined the prognosis of these patients according to the expression of 5-FU related genes. The median of thymidylate synthase (TS), dehydropyrimidine dehydrogenase (DPD), thymidine phosphorylase (TP) and orotate phosphoribosyltransferase (OPRT) mRNA values in the 220 specimens were determined for cut-off levels separating low and high gene expression. In 66 patients, 5-year disease-free survival (DFS) in the TP-high group (n=28) was significantly better than that in the TP-low group (n=38) (P=0.016). Of 220 patients, the 69 patients in TP-high group comprised 28 patients treated with oral fluorouracil derivatives and 41 patients with hormone therapy alone. The proportion of patients with lymph node involvement in the fluorouracil group was significantly greater than that in the hormone therapy alone group (P=0.003). Five-year DFS was not significantly different between the groups (P=0.80). Our results suggest that adjuvant oral fluorouracil chemotherapy may improve the prognosis of the patients with TP high expression breast cancer, and TP mRNA level in breast cancer may predict the effect of new oral fluorouracil derivatives for postoperative adjuvant chemotherapy.

Health-related quality of life in patients with colorectal cancer who receive oral uracil and tegafur plus leucovorin.

OBJECTIVE: Adjuvant chemotherapy with oral uracil/tegafur plus leucovorin has been acknowledged to be a standard treatment for Stage II or III cancer of the colon. The objective of the study was to examine the health-related quality of life during treatment in patients with colorectal cancer who receive oral uracil/tegafur plus leucovorin. METHODS: Health-related quality of life was assessed at baseline (pre-treatment) and at 5-week intervals during treatment, using the European Organization for Research and Treatment of Cancer QLQ-C30 questionnaires. Health-related quality of life data for five courses of treatment were then analyzed longitudinally. RESULTS: Ninety-four patients completed the baseline and post-treatment health-related quality of life assessments. The post-treatment assessments changed significantly from the baseline values and favored post-treatment for all the scales except cognitive function, dyspnea, insomnia, constipation and diarrhea. Role function and social function changed by 10 or more points considered clinically significant. Most of the scales in patients with Grade 0-1 toxicities were better than those with Grade 2-3 toxicities, but Grade 2-3 toxicities were not associated with post-treatment deteriorations in health-related quality of life. The development of Grade 3 toxicities negatively affected on the four scales at the next assessment, compared with Grade 1-2 toxicities. CONCLUSIONS: Overall health-related quality of life did not deteriorate during adjuvant chemotherapy with oral uracil/tegafur plus leucovorin in patients with colorectal cancer, despite the effect from surgical damage, whereas the development of Grade 3 toxicities negatively affected on short-term health-related quality of life. Further comparative studies are needed.

Influence of premature rupture of membrane on the cerebral blood flow in low-birth-weight infant after the delivery.

Cerebral white matter injury, usually called periventricular leukomalacia (PVL), is the most common form of injury to preterm infants that is associated with adverse motor and cognitive outcomes. Intrauterine infection may be an important etiological factor in PVL, and premature rupture of the membranes (PROM) can be identified antepartum. In order to investigate the pathophysiology of cerebral white matter injury induced by PROM, the cerebral blood flow (CBF) of the internal carotid artery and the vertebral artery was measured by neck ultrasonography. The CBF was determined in 84 low-birth-weight infants with gestational ages ranging from 24 to 35 weeks, including 71 infants without PROM and 13 infants with PROM. The mean blood flow velocity and diameter of each vessel were measured on postnatal days 0-70. The intravascular flow volume was determined by calculating the mean blood flow velocity and the cross-sectional area. The mean blood pressures were recorded, and the ejection fraction was determined. The total cerebral blood flow (CBF) was significantly lower in infants with PROM than in infants without PROM from day 10 to day 70. The ejection fraction was significantly higher in infants with PROM than in infants without PROM on days 0, 5, 10, 21, and 42. There was no difference in the mean blood pressure between infants with PROM and infants without PROM. The results of the present study suggest that PROM may decrease cerebral blood flow after the birth.

[Evaluation of 5-fluorouracil-related genes in breast cancer to predict the effect of adjuvant therapy with CMF].

The correlations between mRNA expressions of 5-fluorouracil(5-FU)-related enzymes; thymidylate synthase(TS), dihydropyrimidine dehydrogenase(DPD), thymidine phosphorylase(TP), and orotate phosphoribosyltransferase (OPRT)in breast cancers, along with disease-free survival(DFS), were investigated in 35 patients treated with postoperative adjuvant chemotherapy using cyclophosphamide, methotrexate and 5-fluorouracil(CMF). The patients treated with CMF were divided into two groups, a lower group(L group)and a higher group(H group), according to the median value of the mRNA expression for each enzyme in 220 breast cancer specimens, which were resected between 1996 and 1998 in our institute. 5-year DFS was not significantly different between TS-L and H group(60% and 80%, p=0.38), DPD-L and H group(57.9% and 86.7%, p=0.088), and TP-L and H group(70% and 73.3%, p=0.89), respectively. 5-year DFS in the OPRT-H group(88.9%)was significantly better than that in the OPRT-L group(50%) (p=0.024). In the OPRT-H group, despite the fact that the proportion of patients with lymph node involvement in the CMF group was significantly higher than that in the postoperative adjuvant hormone therapy group, 5-year DFS was not significantly different between the two groups(p=0.10). Our results suggest that OPRT level was the significant predictive marker for DFS in the breast cancer patients treated with postoperative adjuvant chemotherapy using CMF.