[A Breast Stromal Sarcoma-Case Report and Literature Review].

A 83-year-old female presented with a painful, palpable mass of the breast, which had been growing rapidly for the past few months. The physical examination revealed a 5×5 cm, elastic, hard, mobile mass with a smooth surface in the right breast. Ultrasound confirmed a 6×4 cm hypoechoic mass with regular borders in the inferior lateral quadrant of the right breast, which required core-needle biopsy to establish diagnosis. The histopathological result reported stromal neoplasm, suggestive of sarcoma or phyllodes tumor. CT scans of the chest and abdomen were negative for metastases. She underwent simple mastectomy and axillary node sampling. The postoperative course was uneventful, and she hoped no adjuvant therapies to be recommended. She is currently receiving outpatient follow-up care. Breast stromal sarcomas are rare and malignant tumors, hence, treatment protocols are not well-established. Early diagnosis of primary stromal sarcomas is essential, as they tend to spread very rapidly and are known to have poor prognosis.

Correction: Ono, S.; Lam, S.; Nagahara, M.; Hoon, D.S.B. Circulating microRNA Biomarkers as Liquid Biopsy for Cancer Patients: Pros and Cons of Current Assays. J. Clin. Med. 2015, 4, 1890-1907.

The authors wish to make the following corrections to this paper [1]:[...].

Clinicopathological relevance of kinesin family member 18A expression in invasive breast cancer.

Recently, kinesin motor proteins have been focused on as targets for cancer therapy. Kinesins are microtubule-based motor proteins that mediate diverse functions within the cell, including the transport of vesicles, organelles, chromosomes and protein complexes, as well as the movement of microtubules. In the current study, the expression of kinesin family member 18A (KIF18A), a member of kinesin superfamily, was investigated in breast cancer using immunohistochemistry, and its effect on breast cancer prognosis was examined. KIF18A expression level was significantly associated with lymph node metastasis (P=0.047). In patients with high levels of KIF18A expression, survival was significantly poorer compared to patients with low levels of KIF18A expression (disease-free survival, P=0.030). Multivariate analysis revealed that venous invasion (hazard ratio, 9.22; 95% confidence interval, 3.90-23.66; P<0.001) and KIF18A expression (hazard ratio, 3.20; 95% confidence interval, 1.34-6.09; P=0.010) were independent predictive factors for lymph node metastasis. KIF18A may be a useful predictive marker for lymph node metastasis in breast cancer, which could facilitate curative adjuvant treatment.

Circulating microRNA Biomarkers as Liquid Biopsy for Cancer Patients: Pros and Cons of Current Assays.

An increasing number of studies have focused on circulating microRNAs (cmiRNA) in cancer patients' blood for their potential as minimally-invasive biomarkers. Studies have reported the utility of assessing specific miRNAs in blood as diagnostic/prognostic biomarkers; however, the methodologies are not validated or standardized across laboratories. Unfortunately, there is often minimum limited overlap in techniques between results reported even in similar type studies on the same cancer. This hampers interpretation and reliability of cmiRNA as potential cancer biomarkers. Blood collection and processing, cmiRNA extractions, quality and quantity control of assays, defined patient population assessment, reproducibility, and reference standards all affect the cmiRNA assay results. To date, there is no reported definitive method to assess cmiRNAs. Therefore, appropriate and reliable methodologies are highly necessary in order for cmiRNAs to be used in regulated clinical diagnostic laboratories. In this review, we summarize the developments made over the past decade towards cmiRNA detection and discuss the pros and cons of the assays.

Clinicopathologic relevance of claudin 5 expression in breast cancer.

OBJECTIVES: Claudins are major adhesion molecules in tight junctions and are strongly expressed in various cancers. We thus investigated the expression of claudin 5, a member of the claudin family, in breast cancer. METHODS: A total of 193 patients with breast cancer were identified based on their pathologic diagnosis. The expression of each claudin 5 was analyzed in the tumor by immunohistochemical staining. Parametric correlations were done between claudin 5 expression and the clinicopathologic findings. RESULTS: Claudin 5 expression in patients with recurrent breast cancer was statistically significantly higher (P = .004). In addition, analysis of the correlation with other clinicopathologic factors showed statistically significant differences with respect to lymphatic invasion (P = .014), venous invasion (P = .048), estrogen receptor status (P = .002), and human epidermal growth factor 2 status (P = .007). Multivariate analysis revealed that claudin 5 expression was an independent predictive factor in the recurrence for relapse-free survival (RFS) (P = .020). Kaplan-Meier analysis showed that the RFS rate was significantly lower in the high claudin 5 expression group (P = .001). CONCLUSIONS: Patients with breast cancer with high claudin 5 expression had a significantly lower RFS rate. Our findings suggest that claudin 5 may be useful as a new biomarker of a risk factor.

Efficiency of fluorodeoxyglucose positron emission tomography/computed tomography to predict prognosis in breast cancer patients received neoadjuvant chemotherapy.

Neoadjuvant chemotherapy (NAC) has become a standard therapy for patients with advanced breast cancer. Pathological complete response (pCR) after NAC is an important prognostic indicator, but some patients with pCR continue to experience recurrence. So new predictive and prognostic markers in addition to pCR are needed following NAC for breast cancer. Fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) can evaluate metastases in the entire body simultaneously, and has several potential advantages over conventional imaging modalities. The purpose of this study was to evaluate whether FDG-PET/CT can determine NAC response and whether FDG-PET/CT can be a new prognostic marker. We imaged 83 breast cancer tumors with FDG-PET/CT, ultrasound (US), and magnetic resonance imaging (MRI) to evaluate NAC efficacy. As we previously analyzed 110 breast cancers with FDG PET/CT, we defined a threshold of >1.7 maximum standardized uptake value (SUVmax) as abnormal fluorodeoxyglucose (FDG) uptake. After NAC, 16 (19.3 %) tumors had a complete response, 54 (65.1 %) had a partial response, 11 (13.3 %) showed stable disease, and 2 (2.4 %) showed progressive disease. One of the two patients with progressive disease had bone metastasis detected by FDG-PET/CT and was not operated on. Remote metastases were evident in 2.4 % of patients after NAC as determined by FDG-PET/CT. Overall, 17 patients had pathological complete response (pCR). The sensitivity of abnormal FDG uptake after NAC for non-pCR was 20.3 % and the specificity was 94.7 %. Patients with abnormal FDG uptake after NAC experienced significantly more recurrences (P = 0.004) and more of them died (P = 0.010). Moreover, the difference in disease-free survival was more significant in the estrogen receptor (ER)-negative group. FDG-PET after NAC may be more effective for predicting prognosis than for evaluating treatment response. This tendency was particularly remarkable in ER-negative breast cancer tumors. FDG-PET/CT is useful for reevaluating surgical applicability after NAC.

Periostin suppression induces decorin secretion leading to reduced breast cancer cell motility and invasion.

The ability of cancer cells to metastasize is dependent on the interactions between their cell-surface molecules and the microenvironment. However, the tumor microenvironment, especially the cancer-associated stroma, is poorly understood. To identify proteins present in the stroma, we focused on phyllodes tumors, rare breast tumors that contain breast stromal cells. We compared the expression of proteins between phyllodes tumor and normal tissues using an iTRAQ-based quantitative proteomic approach. Decorin was expressed at reduced levels in phyllodes tumor tissues, whereas periostin was upregulated; this result was validated by immunohistochemical analysis of phyllodes tumors from 35 patients. Additionally, by immunoprecipitation and mass spectrometry, we confirmed that decorin forms a complex with periostin in both phyllodes tumors and BT-20 breast cancer cells. Following siRNA-mediated knockdown of periostin in T-47D cells, secreted decorin in the culture medium could be detected by multiple reaction monitoring (MRM). Furthermore, periostin knockdown in BT-20 cells and overexpression of decorin in MDA-MB-231 cells inhibited cell motility and invasion. Our results reveal the molecular details of the periostin-decorin complex in both phyllodes tumor tissues and breast cancer cells; this interaction may represent a novel target for anti-cancer therapy.

[A case of T4bN3c breast cancer treated with modified radical mastectomy with immediate reconstruction using a tissue expander after systemic chemotherapy].

A 40 year-old woman had right T4bN3c breast cancer without distant metastasis. A partial response was achieved in the primary lesion and right supraclavicular and axillary lymph node involvement was no longer noted after systemic chemotherapy (weekly paclitaxel followed by 5-fluorouracil, epirubicin and cyclophosphamide [FEC]). The patient wished to undergo immediate reconstruction of the right breast, and therefore, modified radical mastectomy with tissue expander reconstruction was performed. Adverse effects due to chemotherapy were of Grade 2. Six months after post-mastectomy radiation therapy, the tissue expander was removed and the right breast was reconstructed using an implant. No complications were noted. Six years have passed since the operation, and no local recurrence or distant metastasis has been noted. In addition, the cosmetic appearance of the right breast is satisfactory.

[A case of recurrent breast cancer with carcinomatous pleurisy responding to bevacizumab and paclitaxel therapy].

A 57-year-old woman who had undergone muscle-preserving radical mastectomy at the age of 38 presented with a 5 cm mass in front of the breastbone. She was diagnosed as having recurrent breast cancer( estrogen receptor[ ER] positive, progesterone receptor[ PgR] positive, human epidermal growth factor receptor[ HER] -2 negative) by core needle biopsy. She received an aromatase inhibitor (AI) and showed partial response. At the age of 54, PgR status became negative. At the age of 55, the recurrent tumor increased in size, and the patient received 50 Gy of radiation therapy for its treatment and AI administration was continued. At the age of 57, tumor marker levels increased and detailed examination revealed that the recurrent tumor had increased in size and carcinomatous pleurisy was noted. Bevacizumab and paclitaxel therapy was initiated. Tumor marker levels decreased and the pleural fluid disappeared in 2 weeks. After 3 courses, positron emission tomography( PET) -computed tomography( CT) showed a reduction in the tumor size and a decrease in fluorodeoxyglucose (FDG) uptake. Bevacizumab and paclitaxel therapy could be effective for the treatment of patients with recurrent breast cancer with a life-threatening status, after hormone therapy failure. Bevacizumab and paclitaxel could be effective as first- line chemotherapy because of its good treatment efficiency.

[A case of cardiac tamponade due to breast cancer treated with weekly paclitaxel].

A 44-year-old woman who underwent surgery for left breast cancer 2 years ago presented with dyspnea. Her pathological stage of breast cancer was T2N1M0, ER (3+), PgR (3+), and HER2: 1+. She was treated with tamoxifen only as adjuvant therapy. Pleural effusion in both lungs and pericardial effusion were detected by computed tomography. We aspirated 1,100 mL of the pleural effusion and 700 mL of the pericardial effusion; the cytologies of both were class V. Because her dyspnea disappeared, she was administered weekly paclitaxel. Her pleural effusion and pericardial fluid have not re- emerged. The therapy of choice for pericardial effusion is local chemotherapy or systemic chemotherapy. Systemic chemotherapy after pericardial drainage was effective for this patient. Key words: Breast cancer.

[A case of breast cancer liver metastases with jaundice responding to trastuzumab monotherapy].

We report the case of a 60-year-old female with liver dysfunction resulting from diffuse liver metastases, which, atypically, had originated from breast cancer. She responded remarkably well to trastuzumab monotherapy. She was referred to our hospital because of left breast cancer with multiple general lymphadenopathies. She presented with jaundice and liver dysfunction without a space-occupying lesion or a dilatation of the intrahepatic bile duct on computed tomography images. A liver biopsy was done to rule out autoimmune hepatitis, primary biliary cirrhosis, or primary sclerosing cholangitis, and histopathological examination showed adenocarcinoma in the lymph duct of the liver. Both the primary breast cancer and the liver metastases were negative for hormone receptor expression(ER-, PR-), but overexpressed HER2(HercepTest 3+). She was diagnosed as invasive ductal carcinoma(T1N3cM1, Stage IV). We started trastuzumab monotherapy, which improved her jaundice and liver dysfunction, and resulted in a decrease in lymph node size.

[A long-surviving case of HER2-positive breast cancer with brain metastasis treated by multidisciplinary therapy].

We present a case of a 55-year-old woman who visited our hospital aware of a lump in her right breast. We diagnosed it as bilateral breast cancer [Rt: ABCDE, T3N1M0, ER (-), PgR (-), HER2: 3+, Stage IIIA; Lt: C, T2N0M0, ER (-), PgR (-), HER2: 1+, Stage IIA]. She underwent NAC with EC followed by docetaxel. After cPR, an operation (Rt Bt+Ax, Lt Bp+Ax) was performed. Liver metastases were identified 9 months after the operation, and she was administered weekly paclitaxel+trastuzumab for 12 courses. After cPR, the treatment was changed to trastuzumab only. Because a cerebellar metastasis appeared in postoperative month 19, she underwent an operation using a gamma-knife. Because a new cerebellar metastasis appeared in postoperative month 26, she underwent another gamma-knife operation. Furthermore, liver metastases were diagnosed as PD, and treatment was changed to vinorelbine and trastuzumab. Because third new cerebellar metastasis appeared in postoperative month 45, she underwent another gamma-knife operation. Lung metastases were identified 59 months after the operation, and the therapy was changed to lapatinib and capecitabine. There was no subsequent growth of metastatic tumors, and good control was obtained.

[A case of metastatic breast cancer with bilateral hydronephrosis effectively treated with capecitabine].

A 54-year-old female was diagnosed with invasive ductal carcinoma (pT3N1M0, Stage IIIA, estrogen receptor positive [ER (+)], progesterone receptor positive [PgR (+)], human epidermal growth factor receptor type 2 [HER2] score 0) and was treated by preoperative chemotherapy with weekly paclitaxel followed by 5-fluorouracil(5-FU) plus epirubicin plus cyclophosphamide regimen(FEC). Partial mastectomy with axillary dissection was performed. The pathological examination disclosed that the tumor was scirrhous carcinoma, and a pathological partial response was achieved by chemotherapy. Multiple bone metastases were detected 18 months after the surgery during treatment with letrozole as adjuvant therapy. Retroperitoneal metastases with hydronephrosis and a lung metastasis were detected 28 months after the surgery, even though exemestane and zoledronate were administrated after detection of the bone metastases. Chemotherapy with capecitabine was started and she recovered from hydronephrosis 4 months after the start of treatment. After 32 months from the first treatment with capecitabine, the patient is presently alive without hydronephrosis due to continued chemotherapy.

Microarray analysis of colorectal cancer stromal tissue reveals upregulation of two oncogenic miRNA clusters.

PURPOSE: Cancer stroma plays an important role in the progression of cancer. Although alterations in miRNA expression have been explored in various kinds of cancers, the expression of miRNAs in cancer stroma has not been explored in detail. EXPERIMENTAL DESIGN: Using a laser microdissection technique, we collected RNA samples specific for epithelium or stroma from 13 colorectal cancer tissues and four normal tissues, and miRNA microarray and gene expression microarray were carried out. The expression status of miRNAs was confirmed by reverse transcriptase PCR. Furthermore, we investigated whether miRNA expression status in stromal tissue could influence the clinicopathologic factors. RESULTS: Oncogenic miRNAs, including two miRNA clusters, miR-17-92a and miR-106b-25 cluster, were upregulated in cancer stromal tissues compared with normal stroma. Gene expression profiles from cDNA microarray analyses of the same stromal tissue samples revealed that putative targets of these miRNA clusters, predicted by Target Scan, such as TGFBR2, SMAD2, and BMP family genes, were significantly downregulated in cancer stromal tissue. Downregulated putative targets were also found to be involved in cytokine interaction and cellular adhesion. Importantly, expression of miR-25 and miR-92a in stromal tissues was associated with a variety of clinicopathologic factors. CONCLUSIONS: Oncogenic miRNAs were highly expressed in cancer stroma. Although further validation is required, the finding that stromal miRNA expression levels were associated with clinicopathologic factors suggests the possibility that miRNAs in cancer stroma are crucially involved in cancer progression.

Lymphovascular invasion of colorectal cancer is correlated to SPARC expression in the tumor stromal microenvironment.

As an integral component of the microenvironment in colorectal cancer (CRC), stromal cells can influence tumor progression. Found in the extracellular matrix of CRC, secreted protein acidic and rich in cysteine (SPARC) is expressed in stromal and CRC cells. While SPARC's influence on CRC is not clear, we hypothesized that epigenetically regulated SPARC expression in the microenvironment stromal cells of CRC can affect primary CRC progression and is influenced by lymphovascular invasion (LVI). Quantitative immunohistochemistry (IHC) analysis of paraffin-embedded (n=72) from 37 LVI-positive and 35 LVI-negative primary CRCs was performed. MassARRAY sequencing was performed to assess the methylation status of the promoter region in 22 LVI-positive and 20 LVI-negative CRC and to identify specific CpG island(s) regulating SPARC expression. SPARC in CRC cells was not correlated with LVI, whereas SPARC in the microenvironment stromal cells was inversely related to LVI (P < 0.0001). There was a direct relationship between LVI and 6 specific CpG site methylation in the SPARC promoter region of stromal cells (P = 0.017) but not in CRC cells. Stromal SPARC expression inversely correlated with VEGF-A expression in CRC (P = 0.003) and positively correlated with HSP27 expression (P = 0.009). The results suggested that the epigenetic regulation of SPARC expression in tumor cells versus stromal cells of CRC is significantly different. Stromal cell SPARC expression is epigenetically influenced by LVI of CRC tumors, and may play a significant role in primary CRC progression.

Kinesin 18A expression: clinical relevance to colorectal cancer progression.

Kif18A, a member of the kinesin superfamily of molecular motor proteins, is a microtubule depolymerase and a key regulator of chromosome congregation. Kif18A's role in cancer progression has not been well defined. Our hypothesis is that Kif18A has a role in the progression of colorectal cancer (CRC). To investigate this expression of Kif18A, mRNA was assessed by quantitative real-time PCR in 113 operative specimens of primary CRC. Kif18A was overexpressed and significantly (p < 0.0001) higher in CRC than in normal colon tissue. Kif18A overexpression in CRC significantly correlated with clinicopathologic factors such as tumor stage (p < 0.0001), lymphatic invasion (p = 0.001), lymph node metastasis (p = 0.01), venous invasion (p = 0.002) and peritoneal dissemination (p = 0.02), suggesting that it has a key role in CRC progression. In multivariate analysis, high Kif18A expression had independent significance for poorer overall survival after resection of CRC (p = 0.037). To demonstrate Kif18A's role in CRC progression, we performed translational and in situ studies. Using in vitro studies on CRC cell lines, we evaluated Kif18A's role in proliferation, migration and invasion. CRC cells transfected with Kif18A cDNA demonstrated significant enhanced migration (p < 0.01) and invasion (p = 0.018) compared to mock-transfected cells. When Kif18A was targeted with specific small interfering RNA, CRC cells had significantly reduced proliferation (p < 0.01), migration (p < 0.01) and invasion (p < 0.05). The in vitro and translational studies demonstrated that Kif18A expression is related to events of metastasis and is a significant factor for CRC progression.

Correlated expression of CD47 and SIRPA in bone marrow and in peripheral blood predicts recurrence in breast cancer patients.

PURPOSE: CD47 plays a variety of roles in intercellular signaling. Herein, we focused on the clinicopathologic significance of CD47 expression in human breast cancer. Our data suggest that the correlation between CD47 and signal regulatory protein α (SIRPA) expression may play a key role in the progression of breast cancer. EXPERIMENTAL DESIGN: Quantitative real-time PCR was used to evaluate CD47 mRNA and SIRPA mRNA expression in bone marrow and in peripheral blood from 738 cases of breast cancer. RESULTS: In patients with high levels of CD47 expression in the bone marrow, survival was significantly poorer compared with patients with low levels of CD47 expression [disease-free survival (DFS), P = 0.0035; overall survival (OS), P = 0.015]. Furthermore, high CD47 expression group in a multivariate analysis showed significance as an independent variable for poorer prognosis in DFS (P = 0.024). In the peripheral blood, however, high CD47 expression in patients was not an independent and significant prognostic factor for DFS and OS in a multivariate analysis. CD47 expression was strongly correlated with SIRPA expression in both the bone marrow (P < 0.0001) and peripheral blood (P < 0.0001) of breast cancer patients. CONCLUSIONS: This is one of the first studies to show that a host factor in bone marrow confers prognostic importance. CD47 is an important biomarker in breast cancer, and functions as a prognostic factor for DFS. Moreover, we suggest that the poor prognosis of breast cancer patients with high expression of CD47 is due to an active CD47/SIRPA signaling pathway in circulating cells.

Comprehensive analysis of the clinical significance of inducing pluripotent stemness-related gene expression in colorectal cancer cells.

BACKGROUND: We previously determined that cancer stem-like cells may influence the susceptibility of colorectal cancer (CRC) cells to chemotherapeutic agents. Although Takahashi and Park identified a set of induced pluripotent stem cell (iPS)-related genes required for normal stem cell maintenance, the precise role of iPS-related gene expression in CRC pathogenesis remains to be determined. The purpose of this study was to clarify the clinical relevance of "stemness"-regulating gene expression in CRC cases. MATERIALS AND METHODS: Cancer cells were excised from tissues of 79 CRC cases by laser microdissection (LMD), and quantitative RT-PCR was used to evaluate expression levels of the iPS-related genes c-MYC, SOX2, OCT3/4, LIN28, KLF4, and NANOG, and to identify any associations between their expression and clinicopathological CRC progression. RESULTS: We found that LIN28 expression is significantly associated with lymph node metastasis (p = 0.018) and Dukes stage (p = 0.0319). SOX2expression is also correlated with lymph node metastasis. Furthermore, the ten cases with Dukes D disease expressed significantly higher levels of SOX2transcript than the other 69 cases (p = 0.0136). In contrast, KLF4 expression was inversely related to Dukes stage. Expression of c-MYC, OCT3/4, and NANOG did not appear to have clinical relevance in CRC cases. CONCLUSION: The present analysis strongly suggests that altered expression of several iPS-related genes plays a role in CRC pathogenesis.