Hiroko Nogi

Detection of MUC1 and Keratin 19 mRNAs in the Bone Marrow by Quantitative RT-PCR Predicts the Risk of Distant Metastasis in Breast Cancer Patients

BackgroundEarly detection of micrometastasis in bone marrow is critical for the prognosis of breast cancer patients. Quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) has been used to detect cancer cells in bone marrow, but its utility as a prognostic factor still remains obscure.Materials and MethodsBone marrow samples were aspirated from the anterosuperior iliac spine of 34 patients, immediately after their surgical procedures had been completed. Control samples were also obtained from 10 healthy adult volunteers. The total RNA was extracted from the mononuclear cells, and the expression levels of β-actin, MUC1 and keratin 19 mRNAs were studied by quantitative RT-PCR. Each mRNA level was scored according to the expression level. The sum of these expression scores was defined as the composite expression score, which was employed as the basis of the evaluation.ResultsThe mean follow-up period was 45 months. Nine patients developed distant metastases, and one developed local recurrence. The 4-year disease relapse rates were 75% (RR= 19.38; 95% CI: 1.94-193.20), 28% (RR=3.64; 95% CI: 0.43-31.18), and 8.3% for patients with composite expression scores of 5/6, 3/4 and 2, respectively. The difference among the three groups was statistically significant (log-rank test:p = 0.0029), and multivariate analysis also found the composite expression score to be an independent prognostic factor.ConclusionsBreast cancer patients who show a high composite expression score in bone marrow have a significantly higher risk of recurrence.

Primary Rhabdomyosarcoma of the Breast in a 13-Year-Old Girl: Report of a Case

We report a case of primary alveolar rhabdomyosarcoma of the breast in a 13-year-old Japanese girl. The patient initially presented with a 13 × 8-cm mass in her left breast, which was diagnosed as alveolar rhabdomyosarcoma after an excisional biopsy. Genetic expression of the tumor revealed t(2;13)(q35–37;q14). She underwent modified radical mastectomy (Bt + Ax) and nine lymph nodes were found to be involved. Systemic examinations showed multiple bone and lung metastases 2 weeks after her operation. Despite chemotherapy with doxorubicin, ifosfamide, and actinomycin D, which resulted in remission for 6 months, she died of the disease 8 months after surgery.

Optimization of Region of Interest Drawing for Quantitative Analysis: Differentiation Between Benign and Malignant Breast Lesions on Contrast-Enhanced Sonography.

This study was performed to evaluate the diagnostic utility of quantitative analysis of benign and malignant breast lesions using contrast‐enhanced sonography.

Long-Term Follow-Up of Node-Negative Breast Cancer Patients Evaluated via Sentinel Node Biopsy After Neoadjuvant Chemotherapy

Micro‐Abstract The purpose of this study was to assess the usefulness of sentinel node biopsy (SNB) after neoadjuvant chemotherapy (NAC) in patients with clinically node‐negative breast cancer. SNB after NAC was as accurate as SNB without NAC. Axillary recurrence‐free survival rates were excellent regardless of whether NAC was performed before SNB. Background: Sentinel node biopsy (SNB) is used to accurately assess axillary lymph node status in patients with node‐negative breast cancer. However, its use after neoadjuvant chemotherapy (NAC) is controversial. We retrospectively assessed the usefulness of SNB after NAC by comparing axillary recurrence rates and other parameters in patients with clinically node‐negative breast cancer who underwent SNB after NAC or without NAC. Patients and Methods: At our hospital, 1179 patients with clinically node‐negative breast cancer underwent SNB from April 2007 to December 2013. The clinicopathological and survival data of patients who underwent SNB after NAC (the NAC group) and those who underwent SNB without NAC (the control group) were compared. Patients with a metastatic sentinel node underwent axillary lymph node dissection. Results: The number of patients in the NAC and control groups was 183 (15.5%) and 996 (84.5%), respectively. At diagnosis, tumors were significantly larger in the NAC group (P < .0001). Sentinel nodes were identified in almost all patients in both groups (99.5% in the NAC group vs. 99.8% in the control group). They were nonmetastatic in 147 (80.8%) patients in the NAC group and 849 (85.5%) patients in the control group. At the median follow‐up time of 51.1 months, 6 patients (0.6%) in the control group had axillary lymph node recurrence compared with no patients in the NAC group. Conclusion: SNB after NAC was as accurate as SNB without NAC in patients with clinically node‐negative breast cancer. Axillary recurrence‐free survival rates were excellent regardless of whether NAC was performed before SNB.

An extracellular matrix molecule, secreted by the epithelial-mesenchymal transition is associated with lymph node metastasis of thyroid papillary carcinoma.

Background: Papillary thyroid carcinoma often has lymph node metastasis, compared with follicular thyroid carcinoma. The study showed that epithelial-mesenchymal transition occurs in carcinoma cells during the first stage of metastasis, where some extracellular matrix molecules are secreted in large quantities. Sialic acid carried by fibronectin as the antigen of the monoclonal antibody (MoAb) JT-95, was detected in 90% of papillary thyroid carcinoma cases, and in a few follicular thyroid carcinomas, in the extracellular matrix of thyroid carcinoma cells. Objectives: The current study was conducted to investigate the association between increasing the number of extracellular matrix molecules, fibronectin, and lymph node metastasis. We also co-cultured a thyroid carcinoma cell line and lymphocyte cell line, with and without MoAb JT-95, in order to investigate the mechanism of cell to cell interaction. Patients and Methods: Immunostaining with JT-95 was performed in 45 papillary thyroid carcinoma cases, and 20 follicular type tumors, to investigate the association between the quantity of fibronectin expression and the frequency of lymph node metastasis. The thyroid carcinoma cell line (SW1736), which secreted fibronectin, and the B cell-lymphoma cell line (Daudi), which held integrin on the cell surface, were co-cultured to observe the adhesion of cells to each other. The SW1736 cell line, pretreated with JT-95, was also co-cultured with the Daudi cell line. Results: There were 39 cases with lymph node metastasis in 59 malignant tumors, and 0 cases in 6 benign follicular type tumors. The staining scores by JT-95 of the 39 tumors with lymph node metastasis were 5+ in eight cases and 6+ in 31 cases. On the other hand, the scores of 20 malignant tumors without lymph node metastasis were < 4+ in all of the cases. In the co-cultured assay, numerous adhesions were observed between the SW1736 and Daudi cells. In contrast, the inhibition of adherences was observed in proportion to the concentrations of JT-95. Conclusions: Increased fibronectin expression in thyroid malignancies is correlated with lymph node metastasis.

The predictive value of PgR and HER-2 for response to primary systemic chemotherapy in inflammatory breast cancer.

BackgroundInflammatory breast cancer (IBC) is the most aggressive form of primary breast cancer. We examined the relationship between clinicopathological factors and clinical response to primary systemic chemotherapy (PSC) and outcome.MethodsTwenty-five patients with IBC were examined. Twelve patients received an anthracycline-based regimen, and 13 patients received an anthracycline-and a taxane-containing regimen as PSC. The expression of hormone receptors and human epidermal growth factor receptor-2 (HER-2) was determined by immunohistochemistry.ResultsThe overall clinical response rate was 64.0%. Clinical response to PSC was higher in patients with progesterone receptor (PgR)-positive (P = 0.01) and HER-2-negative (P = 0.03) tumors. Patients with fewer than ten involved axillary lymph nodes (P = 0.01 and P = 0.02, respectively) and with a clinical response to PSC (P = 0.02 and P = 0.01, respectively) showed better distant disease-free survival and overall survival.ConclusionIn patients with IBC, PgR-positive and HER-2-negative tumors are more sensitive to anthracycline-based PSC. Patients with extensive residual tumor (ten or more lymph-nodes involved, no response to PSC) after PSC had unfavorable prognoses.

Abstract P5-07-07: DYRK2 contributes to the generation of breast cancer stem cells through KLF4

Cancer stem cells (CSCs) have been defined by the potential to self-renew and to differentiate. CSCs pose a major hurdle in the treatment of cancer. However, the mechanisms by which cells acquire CSC properties such as drug resistance remain unclear. Dual-specificity tyrosine-regulated kinase 2 (DYRK2) is a protein kinase that phosphorylates its substrates on serine/threonine. Initially, we found that DYRK2 phosphorylates p53 at Ser 46 to regulate apoptotic cell death in response to DNA damage. Recently, we have shown that DYRK2 controls Snail degradation in breast cancer and ovarian serous adenocarcinoma. We also found that knockdown of DYRK2 in luminal-type breast cancer MCF-7 cells increased the cancer stem cell population. Kruppel-like factor 4 (KLF4) is one of the Yamanaka factors. It has been reported that pluripotent stem cells from mouse embryonic or adult fibroblasts are induced by introducing four factors, Oct3/4, Sox2, c-Myc, and Klf4. This finding led us to determine if KLF4 is indispensable for the maintenance of CSCs. The aim of this study is to clarify whether DYRK2 regulates CSCs through KLF4 in breast cancer. Cell lines: MCF-7 (human mammary carcinoma: ATCC) cells were grown according to standard protocols. We established stable DYRK2-depleted cells. MCF-7 cells were transfected with pSuper-puro vector (pSuper control) or pSuper-puro DYRK2 shRNAs (shDYRK2) with puromycin to isolate stable cell lines. In turn, we established both stable DYRK2- and KLF4-depleted cells. shDYRK2 cells were transfected with pSuper-neo vector (pSuper-neo control) or pSuper-neo KLF4 shRNAs (shKLF4) with puromycin and G418. Knockdown of DYRK2 or KLF4 was confirmed by real-time RT-PCR and immunoblotting. The depleted cells were compared with the control cells using real-time RT-PCR, immunoblotting, flow cytometric analysis, mammosphere assay, xenograft models and immunohistological staining. We analyzed the population of breast cancer stem cells by flow cytometric analysis and in vitro mammosphere assay. The results showed that knockdown of DYRK2 was associated with the increase of CD44+/CD24- cells. While pSuper control cells formed mammospheres, they did in a lesser extent compared to shDYRK2 cells. In real-time RT-PCR and immunoblotting analysis, stable DYRK2 depletion in MCF-7 cells induced KLF4 accumulation. We then investigated the effect of KLF4 on stemness by flow cytometric analysis and in vitro mammosphere assay. The results showed that knockdown of KLF4 in shDYRK2 cells reduced the proportion of CD44+/CD24- cells. Whereas shDYRK2/shKLF4 cells formed mammospheres, they did in a lesser extent compared to shDYRK2/pSuper-neo control cells. Moreover, the scale of the mammospheres formed in shDYRK2/shKLF4 cells was significantly smaller, as compared with that in shDYRK2/pSuper-neo control cells. In xenograft models, the loss of KLF4 protein expression significantly decreased tumor formation. Immunohistological staining of fifty-nine samples from surgically treated breast cancer patients showed an inverse correlation between DYRK2 and KLF4 expression. These findings revealed that DYRK2 contributes to the generation of breast cancer stem cells through KLF4. Citation Format: Imawari Y, Mimoto RK, Yamaguchi N, Kamio M, Kato K, Nogi H, Toriumi Y, Uchida K, Takeyama H, Yoshida K. DYRK2 contributes to the generation of breast cancer stem cells through KLF4 [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P5-07-07.

Nonrecurrent inferior laryngeal nerves and anatomical findings during thyroid surgery: report of three cases

A nonrecurrent inferior laryngeal nerve (NRILN) is found more frequently on the right side than on the left, and it is closely associated with an aberrant right subclavian artery. The presence of the aberrant right subclavian artery on preoperative computed tomography (CT) scan suggests NRILN; however, different types of branching locations and pathways exist. Here, we report three NRILN cases with different pathways where the vagus nerve arises more medial than usual and a review of the literature. Case 1: A 30-year-old Japanese female presented with papillary thyroid carcinoma. Preoperative CT scan revealed an aberrant right subclavian artery, and an operation was performed under suspicion of NRILN. During the operation, the vagus nerve was found to arise more medially than usual and two NRILNs originated from it at the level of the cricoid cartilage and at a more caudal position; the two NRILNs were preserved. Case 2: A 33-year-old Japanese female with a thyroid nodule of increased size underwent surgery. Preoperative CT scan revealed an aberrant right subclavian artery, which suggested NRILN. During the operation, the vagus nerve was identified to run more medially than usual and NRILN was found to originate at the level of the cricoid cartilage; NRILN was preserved. Case 3: A 78-year-old Japanese female underwent an operation with a diagnosis of papillary thyroid carcinoma. Preoperative CT scan showed an aberrant right subclavian artery. During the operation, NRILN was found to originate from the vagus nerve at the level of the lower pole of the thyroid gland, and the vagus nerve ran medial to the common carotid artery at the caudal level.

Abstract P2-12-14: The effect of low intensity exercise for mental status and quality of life of Japanese early breast cancer patients

Objective: Breast cancer is one of the most common cancers, and it accounts for 20% of malignant tumors of women. Psychological damage of the breast cancer patient is serious. In our institution, approximately 30% of the early breast cancer patients diagnosed as depressed mental status before operation. Breast cancer patients have to receive hormone therapy, chemotherapy, and radiation therapy for a long term after operation. Depressed mental status affects execution of the treatment, and the interruption of the treatment brings the risk of cancer recurrence. As a treatment for depressed mental status of cancer patients, medication and psychotherapy are common, but some previous reports showed exercise is effective. Exercises provided in these reports were various kinds and strength, and the standard prescription is not established. We investigate the effect of walking program as a mild aerobic exercise, whether it improves the depressed mental status and quality of life (QOL) of breast cancer patients. Methods: 25 early breast cancer patients were recruited. Depressed mental status and QOL were assessed by Center for Epidemiologic Studies Depression Scale (CES-D) and Medical Outcomes Study 36-item Short Form Health Survey (SF-36). Physical activity was measured by accelerometer (Lifecorder PLUS, Suzuken), and estimated using metabolic equivalent foe task (MET). Participants mounted accelerometer after discharge, and baseline physical activity was recorded till 1 month after operation. After recording baseline physical activity, we instructed to perform walking or mild aerobic exercise. Scores of CES-D and SF-36, and physical activity data were measured again after two months. Results: Physical activity after intervention was significantly increased as compared to baseline (8.5±5.5 vs 12.3±6.9 MET.h/week, p=0.02 ). At pre-operation phase, nine patients (36%) regarded as depressed mental status by CES-D, and depressed patients decreased to five at the time of 3 months after operation. A subscale of SF-36 ( mental health ) reduced significantly compared with pre-operation and 3 months after operation ( p Conclusion: This study demonstrated the possibility that walking program as a mild exercise improves mental status and QOL of perioperative breast cancer patients. It is necessary to examine in more detail for clinical application. Citation Format: Makiko Kamio, Kazumi Kawase, Hiroko Nogi, Ken Uchida, Hiroshi Takeyama. The effect of low intensity exercise for mental status and quality of life of Japanese early breast cancer patients [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-12-14.

Abstract P2-06-14: Clinical relevance and biological properties of oligometastatic breast cancer in lung; prognostic impact of CD44+/CD24−/low cells

Background: Metastatic breast cancer is a systemic disease. Our aim was to evaluate the clinical outcomes of pulmonary metastasectomy of recurrent breast tumors and to identify possible prognostic factors. Methods: We reviewed data from a registry of patients with lung metastases from breast tumors who received pulmonary metastasectomy in Jikei University Hospital between 2004 and 2011. We analyzed prognostic factors for overall survival (OS) and progression free survival (PFS) after metastasectomy. We also investigated lung metastases for the prevalence of CD44+/CD24−/low tumor cells and evaluated their prognostic significance. Results: Among 17 patients with lung metastasis of breast tumors, 5-year OS and PFS were 72% and 36%, respectively. Better OS was observed among patients with oligometastatic breast cancer (OMBC). Patients with OMBC, estrogen receptor (ER) positive cells, and disease free intervals (DFI) of >8 years had better PFS. The average prevalence of CD44+/CD24−/low tumor cells in lung metastases of breast cancer was 21%, ranging from 0 to 90%. The presence of CD44+/CD24−/low tumor cells influenced the progression after lung metastasectomy, with median PFS times of only 6 months in patients with high-prevalence of cancer-initiating cells. CD44+/CD24−/low cells with cancer-initiating properties were present in only 9% ± 12 of patients with OMBC but were found in 73% ± 21 of patients with non-OMBC. Conclusion: Pulmonary metastasectomy may be a treatment option for OMBC patients with lung metastases. Better prognosis of OMBC may be related to low levels of cancer-initiating cells. Citation Format: Rei Mimoto, Tadashi Kobayashi, Yoshimi Imawari, Makiko Kamio, Kumiko Kato, Hiroko Nogi, Yasuo Toriumi, Ken Uchida, Hiroshi Takeyama. Clinical relevance and biological properties of oligometastatic breast cancer in lung; prognostic impact of CD44+/CD24−/low cells [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-06-14.