Akemi Kataoka

The clinical significance of MMP-1 expression in oesophageal carcinoma

Matrix metalloproteinase-1 (MMP-1) is involved in the degradation of interstitial collagen and thus thought to play a role in invasion of carcinoma. We investigated 51 oesophageal carcinoma patients to clarify the significance of MMP-1. MMP-1 mRNA was demonstrated to be expressed exclusively in almost all carcinoma tissue specimens (T) (94.1%) by reverse transcription-polymerase chain reaction, but not found in normal mucosal tissue specimens (N). The mean T/N ratio of MMP-1 was 42.5 and cases with T/N ≥ 10 had a higher incidence of cases involving muscularis propria than those with T/N < 10 which included all the cases involving the submucosa (P< 0.05). MMP-1 mRNA was significantly associated with both 40 kD (putative active MMP-1) and 50 kD (putative latent MMP-1) gelatinolytic bands (n=17). These findings indicated that MMP-1 mRNA reflected the net function of MMP-1 and suggested MMP-1 to be involved in carcinoma invasive process. On the other hand, MMP-1 mRNA was inversely correlated with the patient prognosis (P< 0.01). These results indicated that MMP-1 might therefore play a crucial role in local invasion, but not in systemic dissemination. As a result, MMP-1 might be a novel prognostic factor independent from those previously reported in oesophageal carcinoma.

Modulation of thymidine phosphorylase by neoadjuvant chemotherapy in primary breast cancer

The combination effect of docetaxel and capecitabine on tumour response rate and survival was demonstrated recently in metastatic breast cancer patients. This combination was based on an experimental hypothesis that taxane can increase tumour sensitivity to the effect of capecitabine through the upregulation of thymidine phosphorylase (TP), which is responsible for the metabolism of 5-fluorouracil (5-FU) and its derivatives, including capecitabine. To examine the alteration in TP expression before and after neoadjuvant chemotherapy, 92 patients with primary breast cancer (T2-4N0-1M0) were enrolled in this study; 14 were treated with adriamycin and cyclophosphamide (AC) or epirubicin and cyclophosphamide (EC); 58 with 5-FU, adriamycin, and cyclophosphamide (FAC) or 5-FU, epirubicin, and cyclophosphamide (FEC); and 20 with FEC followed by docetaxel/taxotere (TXT-containing regimen). Thymidine phosphorylase upregulation was seen in 54.4% and 32.6% of patients in tumour cells and stromal cells, respectively. Increases in TP expression were found only in the AC/EC and TXT-containing regimen groups. In conclusion, it was strongly suggested that unlike 5-FU-containing regimens, the taxane and AC combination therapies upregulate TP expression in primary breast cancer. Thymidine phosphorylase upregulation by several anticancer drugs implies the importance of individualised strategies for sensitisation of tumour tissues to 5-FU and its derivatives.

Frequency of Microsatellite Instability inBreast Cancer Determined by High-Resolution Fluorescent Microsatellite Analysis

In breast cancer, the rates of positivity for microsatellite instability (MSI), vary greatly in the literature. Using high-resolution fluorescent microsatellite analysis (HFRMA), we studied microsatellite alterations in 75 patients with sporadic breast cancer. In this system, several devices were prepared to improve reproducibility of polymerase chain reaction products, migration accuracy of electrophoresis, and characteristics of the detection system. Precise and objective analyses of microsatellite alterations are made feasible using HRFMA. Seven of the 75 cases were judged to be positive for MSI, the rate of positivity being 9.3%. This rate is relatively low compared to the data in the literature. All the microsatellite changes observed in this system can be classified into two types: type A with relatively small changes in microsatellite sequences observed in limited loci and type B with drastic and widely dispersed changes. The former was thought to be connected to abnormal activity in DNA mismatch repair (MMR). Among the 7 cases, 6 (8.0%) had type A alterations, which means that the tumors may have an abnormal MMR activity. Application of precise and objective systems for microsatellite analysis is expected to be clinically useful to detect patients at high risk for cancers.

Identification of recurrence-related microRNAs in the bone marrow of breast cancer patients

Recently, bone marrow has been considered as playing a critical role in the generation of both metastasis and recurrent disease. The accumulation of a single microRNA in the bone marrow has the potential to regulate the translation of multiple genes in cancer metastasis and may therefore serve as a prognostic marker for cancer recurrence. MicroRNA microarray analysis was performed to compare microRNA levels in bone marrow from 4 breast cancer patients with recurrent disease and 4 patients without recurrence. Accumulation of two of these microRNAs, miR-21 and miR-181a, in the recurrent breast cancer cases was validated by RT-PCR in bone marrow from 291 additional breast cancer cases. Expression of a common target gene, PDCD4, was also determined in bone marrow from 291 breast cancer cases. Increased miR-21 and miR-181a levels were significantly associated with shortened disease-free survival (DFS; p=0.0003, 0.0007) and overall survival (OS; p=0.0351, 0.0443), respectively. While low PDCD4 expression was also significantly associated with poorer DFS (p=0.036). Multivariate analysis identified bone marrow miR-21 and mR-181a levels as valuable independent prognostic factors, with correlation coefficients that were significantly higher than that of the transcript of their common target gene. Accumulation of miR-21 and miR-181a in bone marrow appears to be associated with prognosis in breast cancer patients. The much higher significant correlation with microRNA levels and prognosis suggests epistatic effects on multiple target genes in the bone marrow of breast cancer patients.

Evaluation of circulating tumor cells in patients with breast cancer: multi- institutional clinical trial in Japan

BackgroundWith the development of the CellSearch System, it has become possible to measure circulating tumor cell (CTC) levels with high reproducibility, and the CTC test is currently being used clinically for patients with metastatic breast cancer in the United States. It is imperative that the clinical significance of the CTC test also be examined in Japan.MethodsUsing the CellSearch System, CTC levels were evaluated in 57 healthy individuals and patients with benign breast disease; 30 patients with primary breast cancer (stages 1–3); and 38 patients with metastatic breast cancer. First, the relationship between CTC levels and the presence of metastasis was examined using a cutoff score of 2 CTCs per 7.5 ml whole blood. Then, the patients with metastatic breast cancer were divided into two groups, using a cutoff score of 5 CTCs per 7.5 ml blood, and progression-free survival (PFS) and overall survival (OS) were compared in the two groups.ResultsWhen the clinical cutoff score was set at 2 CTCs per 7.5 ml blood, 0% of the healthy individuals and patients with benign breast disease (0/57), 3.3% of the patients with primary breast cancer (1/30), and 50% of the patients with metastatic breast cancer (19/38) were identified as as having 2 CTCs per 7.5 ml blood. Additionally, with a cutoff score of 5 CTCs, 11 patients were reported to have 5 or more CTCs and both PFS (P = 0.0036) and OS (P = 0.04) were worse for this patient population than for the population with fewer than 5 CTCs.ConclusionAs concluded in a similar clinical trial in the United States, for patients with breast cancer, measuring CTC levels can be both an accurate indicator of metastases and an important measure of patient prognosis.

Prognostic significance of the coexpression of p53 protein and c-erbB2 in breast cancer.

BACKGROUND The expressions of p53 protein and c-erbB2 have been thoroughly analyzed as prognostic factors for breast cancer. However, the prognostic significance of the coexpression of p53 protein and c-erbB2 remains controversial. METHODS The immunohistochemical expression of p53 protein and c-erbB2 was evaluated in 242 women with breast cancer. RESULTS According to the combination of p53 protein and c-erbB2, a group negative for both (125 cases), a group positive for either one of these two parameters (99 cases) and a group positive for both (18 cases) were identified. The group positive for one factor had a significantly (P = 0.0045) worse disease-free survival (DFS) than the group negative for both, while the group positive for both had a significantly (P = 0.0023) worse DFS than the group positive for one factor. A multivariate analysis indicated that the relative risk of p53 protein alone and cerbB2 alone was 2.18 and 2.93, respectively, while the relative risk of the group positive for one factor and the group positive for both was 2.29 and 6.37, respectively . CONCLUSIONS Breast cancer with a coexpression of p53 protein and c-erbB2 was thus found to have a more significant prognostic value than those with a positive expression for either of these two biological parameters, while the prognostic significance of these two parameters themselves remained independent of each other.

Early experience of endoscopic extirpation of benign breast tumors via an extra-mammary incision

The early results of 6 patients with a benign breast mass who underwent an endoscopic extirpation via an extra-mammary incision were presented. Under general anesthesia in either a lateral or supine position, a 12-mm and two 5-mm incisions in the infra-mammary line in 2 patients and in the mid-axillary line in the remaining 4 patients were made. The tumors were then endoscopically extirpated. Preoperative aspiration needle cytology revealed six fibroadenomas in 5 patients and one intraductal papilloma in the other patient. All patients were single females with a mean age of 22.5 years. The maximal size of the masses was 5 cm on average. The average operation time was 3 hours 20 minutes. Regarding postoperative complications, subcutaneous emphysema extending to the neck due to CO2 gas inflation and a burn in the skin were seen in 1 patient each; however, no further treatment was required in these cases. The postoperative hospital stay was 1.7 days on average, and all patients were extremely satisfied with the cosmetic results of the procedure. The cosmetic results are drastically improved by the application of endoscopic removal via extra-mammary approaches, which are newly introduced, for benign breast tumors.

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. Clin Cancer Res; 16(18); 4625–35. ©2010 AACR.

Comparison of the Immunohistochemical Expression of EGFR, c-erbB2 and p53 Protein between Primary and Recurrent Breast Cancer

BackgroundAlthough various biological parameters have been evaluated as predictors of the response to chemohormonal therapy for breast cancer, few studies have comparied the biological parameters of the primary and recurrent breast cancers.Patients and MethodsThe immunohistochemical expression of epidermal growth factor receptor (EGFR), c-erbB2 and p53 protein were analyzed on both primary and matching recurrent lesions from 42 patients with breast cancer.ResultsEGFR and c-erbB2 expression were concordant between the primary and matching recurrent lesion in 27 (90%) of 30 cases in which EGFR was evaluated and in all (100%) 12 cases in which c-erbB2 was evaluated. Twelve (67%) of 18 cases in which p53 protein was evaluated showed concordance of p53 protein expression between the primary and recurrent lesions, while 6 other cases did not. The intensity of the immunoreactivity of p53 protein was 10-50% staining in 5 of these 6 cases, while only one case showed greater than 50% staining of p53 protein in the primary lesion and negative staining in the matching recurrent lesion.ConclusionEGFR and c-erbB2 immunoreactivity were concordant between the primary and matching recurrent lesions in the majority of the breast cancer cases. In addition, some cases were not concordant regarding the intensity of immunoreactivity for p53 protein, whereas few cases showed both a strong positivity and negative finding for p53 protein between the primary and matching recurrent lesions.

Prognostic Significance of the Combination of Biological Parameters in Breast Cancer

Abstract.To evaluate whether the combination of biological parameters increases their prognostic value, the expression of epidermal growth factor receptor (EGFR), DNA ploidy, and estrogen receptor (ER) status were analyzed on 998 patients with breast cancer. Poor findings for each biological parameter were positive for EGFR, aneuploid for DNA ploidy, and negative for ER. According to the number of poor findings in these three parameters, the groups with none (309 cases), one (377 cases), two (161 cases), and three (151 cases) poor findings were classified. A significant (P < 0.0001) difference was found in the disease-free survival (DFS) among the four groups. A multivariate analysis indicated the combination of three biological parameters to be an independently significant factor for DFS, while the relative risk gradually increased as the number of poor findings increased. In conclusion, the present study indicated a gradual increase in the prognostic significance as the number of combined biological parameters increased.