Increased centrosome number in BRCA-related breast cancer specimens determined by immunofluorescence analysis.

BRCA-related breast carcinoma can be prevented through prophylactic surgery and an intensive follow-up regimen. However, BRCA genetic tests cannot be routinely performed, and some BRCA mutations could not be defined as deleterious mutations or normal variants. Therefore, an easy functional assay of BRCA will be useful to evaluate BRCA status. As it has been reported that BRCA functions in the regulation of centrosome number, we focused on centrosome number in cancer tissues. Here, 70 breast cancer specimens with known BRCA status were analyzed using immunofluorescence of γ-tubulin (a marker of centrosome) foci. The number of foci per cell was higher in cases with BRCA mutation compared to wild-type cases, that is, 1.9 (95% confidence interval [CI], 1.5-2.3) vs 0.5 (95% CI, 0.2-0.8) (P < .001). Specifically, foci numbers per cell in BRCA1 and BRCA2 mutation cases were 1.2 (95% CI, 0.6-1.8) and 2.2 (95% CI, 1.7-2.6), respectively, both higher than those in wild-type cases (P = .042 and P < .0001, respectively). The predictive value of γ-tubulin foci as determined by area under the curve (AUC = 0.86) for BRCA status was superior to BRCAPRO (AUC = 0.69), Myriad Table (AUC = 0.61), and KOHBRA BRCA risk calculator (AUC = 0.65) pretest values. The use of γ-tubulin foci to predict BRCA status had sensitivity = 83% (19/23), specificity = 89% (42/47), and positive predictive value = 77% (20/26). Thus, γ-tubulin immunofluorescence, a functional assessment of BRCA, can be used as a new prospective test of BRCA status.

Quantitative diagnostic imaging of cancer tissues by using phosphor-integrated dots with ultra-high brightness.

The quantitative sensitivity and dynamic range of conventional immunohistochemistry (IHC) with 3,3'-diaminobenzidine (IHC-DAB) used in pathological diagnosis in hospitals are poor, because enzyme activity can affect the IHC-DAB chromogenic reaction. Although fluorescent IHC can effectively increase the quantitative sensitivity of conventional IHC, tissue autofluorescence interferes with the sensitivity. Here, we created new fluorescent nanoparticles called phosphor-integrated dots (PIDs). PIDs have 100-fold greater brightness and a more than 300-fold greater dynamic range than those of commercially available fluorescent nanoparticles, quantum dots, whose fluorescence intensity is comparable to tissue autofluorescence. Additionally, a newly developed image-processing method enabled the calculation of the PID particle number in the obtained image. To quantify the sensitivity of IHC using PIDs (IHC-PIDs), the IHC-PIDs method was compared with fluorescence-activated cell sorting (FACS), a method well suited for evaluating total protein amount, and the two values exhibited strong correlation (R = 0.94). We next applied IHC-PIDs to categorize the response to molecular target-based drug therapy in breast cancer patients. The results suggested that the PID particle number estimated by IHC-PIDs of breast cancer tissues obtained from biopsy before chemotherapy can provide a score for predicting the therapeutic effect of the human epidermal growth factor receptor 2-targeted drug trastuzumab.

Minimal impact of postmastectomy radiation therapy on locoregional recurrence for breast cancer patients with 1 to 3 positive lymph nodes in the modern treatment era.

INTRODUCTION: Given modern treatment strategies, controversy remains regarding whether postmastectomy radiation therapy (PMRT) is necessary for breast cancer patients with 1-3 positive axillary lymph nodes (ALN). Our aim was to assess the significance of PMRT in the modern treatment era for these patients. MATERIAL AND METHODS: We have conducted the retrospective multicenter study and identified 658 patients with 1-3 positive ALN who were treated with mastectomy and ALN dissection between 1999 and 2012. Propensity score weighting was used to minimize the influence of confounding factors between the PMRT and no-PMRT groups. The variables including tumor size, lymph nodes status, skin and/or muscle invasion, histological grade, lymphovascular invasion and ER positivity which were statistically unbalanced between the groups were used to define the propensity scores. RESULTS: The median follow-up time was 7.3 years. In the modern era (2006-2012), no significant difference in locoregional recurrence (LRR)-free survival was noted between the PMRT and no-PMRT groups (P = 0.3625). The 8-year LRR-free survival rates of the PMRT and no-PMRT groups were 98.2% and 95.3%, respectively. After matching patients by propensity scores, the PMRT group, compared to the no-PMRT group, exhibited significantly better locoregional control (P = 0.0366) in the entire cohort. The 10-year LRR-free survival rates were 97.8% and 88.4% in the PMRT and no-PMRT groups, respectively. In contrast, no significant difference in LRR-free survival was noted between the PMRT and no-PMRT groups in the modern era (P = 0.5298). The 8-year LRR-free survival rates of patients treated in the modern era were approximately the same between the groups (98.0% and 95.7% in the PMRT and no-PMRT groups, respectively). Particularly, LRR-free survival of HER2 positive breast cancer significantly improved in the modern treatment era, compared with that of the old treatment era (P = 0.0349). CONCLUSION: PMRT had minimal impact on LRR for breast cancer patients with 1-3 positive ALN in the modern treatment era.

Combined Immunohistochemistry of PLK1, p21, and p53 for Predicting TP53 Status: An Independent Prognostic Factor of Breast Cancer.

It is difficult to predict the TP53 status by p53 immunohistochemistry (IHC). We aimed to improve the accuracy of p53 IHC with p53-regulated proteins for predicting the TP53 mutation status. TP53 mutations were detected in 19 of 38 breast cancer patients (50%). Five of 7 cases of protein-truncating mutation of TP53 were completely negative for p53 IHC, whereas 11 of 12 cases of TP53 point mutation were strongly positive for p53 IHC. Therefore, to avoid false negatives, we extracted p53-dependent universally downregulated genes using microarray analysis from 38 breast cancer patients and 2 p53-inducible cell lines. From 9 commonly repressed genes, we evaluated 3 genes, baculoviral IAP repeat-containing 5 (BIRC5), polo-like kinase 1 (PLK1), and BUB1 mitotic checkpoint serine/threonine kinase (BUB1), which were previously identified as p53-dependent repressed genes. PLK1≥Allred total score (TS) 5 showed the highest correlation with TP53 mutation. To decrease false positivity, we evaluated p21 IHC. Although strong staining of p21 was observed in 4 cases (10.5%), all 4 were wild-type TP53. Thus, p53 mutation-like (p53mt-like) IHC was identified by p53 TS7,8 with PLK1≥TS 5 and p21 TS≤6. p53 mt-like IHC correlated with TP53 mutation (predictive value=0.94). In other 157 breast cancer cases, p53 mt-like was an independent prognostic marker in multivariate analysis and a strong prognostic factor. Stratification with p53 mt-like IHC identified patients with a poorer prognosis. In conclusion, we identified reliable IHC conditions to predict the TP53 status of breast cancer patients.

[A case of advanced gastric and rectal cancer (double cancer) successfully treated with mFOLFOX6 therapy].

A 75-year-old man was diagnosed with gastric cancer (UL post c0- II c (c T1N0) and M-less ctype II (cT2N0)) and rectal cancer (Rb ctype II (cT2N1) with multiple lung metastases (M1). The patient was treated with modified (m) FOLFOX6 regimen (oxaliplatin in combination with infusional 5-fluorouracil/Leucovorin). Chest and abdominal CT scan revealed that multiple lung metastases and abdominal lymph node metastases were obviously reduced in size. The primary lesion of the rectum almost disappeared on endoscopic examination. As for the lesions of the stomach, the UL post c0- II c lesion completely disappeared, and the M-less ctype II lesion was reduced remarkably. Thus, a significant reduction of the tumors was observed. This case suggests that mFOLFOX6 regimen can be an option for gastric cancer.

Assessing the extent of breast cancer using multidetector row helical computed tomography.

OBJECTIVE: The aim of the study was to evaluate the usefulness of multidetector row helical computed tomography (MD-CT) in assessing the local extent of breast cancer. METHODS: Seventy-five breast cancer patients were examined using MD-CT with scanning performed in the supine position at 1-mm collimation. The extent of the breast tumors determined using CT was compared with that based on histopathologic mapping with continuous 5-mm slices. RESULTS: The CT evaluation of the maximum diameter of the extent of breast cancer was much better correlated with the histopathologic diagnosis (correlation coefficient=0.90) than the pre-CT diagnosis (correlation coefficient=0.46). Computed tomography correctly detected mammographically and clinically occult cancer other than the index lesion in 14 of 15 patients. The sensitivity, specificity, and accuracy in the diagnosis of the additional lesions were 93.3%, 98.3%, and 97.3%, respectively. Because the images were obtained in the supine position, they were useful for surgical planning. CONCLUSION: The extent of breast cancer can be determined accurately using MD-CT.

Pathological assessment of intraductal spread of carcinoma in relation to surgical margin state in breast-conserving surgery.

BACKGROUND: Spreading of carcinoma has been considered to be a prognostic factor for local failure after breast-conserving therapy. The extensive intraductal component (EIC) was defined as when the component of intraductal carcinoma constitutes more than 25% of the primary tumor with intraductal foci. However, the definition of EIC was based on the predominance of intraductal component surrounding the invasive lesions and not on the segmental anatomy. We designated carcinoma extension as the intraductal spread of carcinoma (ISC) along with the duct-lobular system by three-dimensional (3-D) reconstruction analysis. This study was initiated to simplify the method of two-dimensional (2-D) pathological examination based on 3-D mapping. METHODS: Thirty-four specimens from breast cancer patients were subjected to 3-D reconstruction. We investigated the correlation between actual extension of intraductal carcinoma and EIC defined by 2-D examination or ISC grading defined by 3-D reconstruction. Furthermore, using another 62 histological mappings, we investigated how correctly the simplified 2-D method using several paraffin blocks reflected the actual carcinoma spread and margin state. RESULTS: Carcinoma extension over 2 cm was observed in 64% specimens that were EIC positive and 26% specimens that were EIC negative. In contrast, according to the ISC grading defined by 3-D reconstruction, none of the specimens with a low grade of ISC demonstrated carcinoma extension over 2 cm. Carcinoma extension over 2 cm was observed in 71% of specimens with a high grade of ISC, thus demonstrating a correlation between carcinoma extension and ISC grading. In addition, the simplified 2-D method using only several blocks reflected both the 3-D ISC grading and surgical margin state. CONCLUSIONS: We conclude that ISC grading correlates with carcinoma extension and surgical margin state. From a clinical point of view, the simplified 2-D examination using paraffin blocks may contribute to routine surgical pathology in evaluating the degree of carcinoma extension in breast-conserving therapy.