Malignant Breast Research Articles

Abstract PR06: Bacteroides fragilis: A potential pathogen orchestrating EMT and stemness in breast epithelial cells via concomitant activation of Notch and βcatenin axes

Abstract Background: The last decade established significant contributions of microbiome to many organ-specific cancers. A few recent studies suggested the existence of distinct breast microbiota and a shift in microbial community composition in diseased breast compared to normal breast; however, their functional impact and underlying mechanisms are unknown. The present study was designed to examine the contribution of procarcinogenic bacteria in breast cancer initiation, growth, and progression. Results: Utilizing extensive data mining and metagenomic analyses, we discovered the presence of toxin-producing Bacteriode fragilis (B. fragilis) in malignant breast. B. fragilis is a procarcinogenic bacteria known for its potential to initiate and/or promote colon cancer, and its pathogenicity has been attributed to its unique toxin “Fragilysin” or “B. fragilis toxin (BFT).” Mice infected with B. fragilis exhibited a significant increase in circulating BFT and distinct morphologic alterations in mammary gland. While no changes were observed in cell growth and clonogenicity upon BFT treatment, significant increase in migration and invasion potential and decreased adhesion of MCF10A and MCF7 cells were observed. BFT-treated cells displayed acquisition of fibroblast-like appearance and increased formation of pseudopodia/microtentacles emanating from the cell membrane along with molecular markers of epithelial-to-mesenchymal transition. Decreased expression of epithelial marker E-cadherin along with elevated levels of mesenchymal markers, N-cadherin, and vimentin were observed. BFT also increased the expression and nuclear translocation of EMT-related transcription factors, Snail, Slug, and Twist. BFT-treated cells attained stem cell-like phenotype exhibiting an increased ability to form secondary and tertiary mammospheres and elevated expression of pluripotency-factors (Oct4, Nanog, and Sox2). Mechanistic studies showed that BFT induced expression and nuclear translocation of NICD (cleaved NOTCH) and βcatenin, resulting in activation of downstream targets. Inhibition of Notch1 and βcatenin using γ-secretase inhibitor and ICG001 successfully inhibited functional effects of BFT. Further, we found that BFT-pretreated MCF7 cells exhibit increased tumor growth and form multifocal tumors in mice. MCF10A-KRas cells, pretreated with BFT, also showed increased tumor progression and multifocal tumors in mice. In vivo limiting dilution assay using breast tumors from BFT-pretreated MCF7 cells exhibited a striking increase in tumor-initiating cells. Follow-up analyses of these tumors demonstrated increased migratory, invasive, and mammospheres-forming behavior, confirming that brief BFT exposure elicits long-term molecular changes. Conclusion: Collectively, these findings present the first in vitro and in vivo evidence to show that Bacteriode fragilis toxin induces EMT, invasion/migration, and stem cell-like phenotype and leads to concomitant activation of Notch and βcatenin axes. This abstract is also being presented as Poster A24. Citation Format: Sheetal Parida, Shaoguang Wu, Nethaji Muniraj, Sumit Siddharth, Arumugam Nagaligam, Christina Hum, Panagiotis Mistriotis, Konstantinos Konstantopoulos, Cynthia Sears, Dipali Sharma. Bacteroides fragilis: A potential pathogen orchestrating EMT and stemness in breast epithelial cells via concomitant activation of Notch and βcatenin axes [abstract]. In: Proceedings of the AACR Special Conference on the Microbiome, Viruses, and Cancer; 2020 Feb 21-24; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2020;80(8 Suppl):Abstract nr PR06.

Abstract PR02: Bacteroides fragilis toxin induces epithelial-to-mesenchymal transition and stem-like phenotype in breast epithelial cells and concomitantly activates Notch1 and βcatenin axes

Abstract The last decade established significant contributions of the microbiome to many organ-specific cancers. A few recent studies suggested the existence of distinct breast microbiota and a shift in microbial community composition in diseased breast compared to normal breast; however, their functional impact and underlying mechanisms are unknown. The present study examines the contribution of procarcinogenic bacteria in breast cancer initiation, growth, and progression. Extensive data mining and metagenomic analyses of existing datasets revealed the presence of toxin-producing Bacteroides fragilis in malignant breast. B. fragilis is a procarcinogenic bacteria known for its potential to initiate and promote colon cancer; its pathogenicity has been attributed to its unique toxin “BFT.” B. fragilis-infected mice exhibited a significant increase in circulating BFT and distinct morphologic alterations in mammary gland. In vitro, upon treatment with BFT, no changes were observed in cell growth and clonogenicity. However, significant increase in migration and invasion potential and decreased adhesion of MCF10A and MCF7 cells were observed. BFT-treated cells displayed acquisition of fibroblast-like appearance and increased formation of pseudopodia/microtentacles emanating from the cell membrane along with molecular markers of epithelial-to-mesenchymal transition. Decreased expression of epithelial marker, E-cadherin, along with elevated levels of mesenchymal markers, N-cadherin and vimentin, were observed. BFT also increased the expression of EMT-related transcription factors, Snail, Slug and Twist. BFT-treated cells attained stem cell-like phenotype, exhibiting an increased ability to form secondary and tertiary mammospheres and elevated expression of pluripotency-factors (Oct4, Nanog, and Sox2). Mechanistic studies showed that BFT induced expression and nuclear translocation of NICD (cleaved NOTCH) and β-catenin resulting in activation of downstream targets. Inhibition of Notch1 and β-catenin using γ-secretase and β-catenin inhibitors successfully inhibited functional effects of BFT. Mammary gland implantation and in vivo limiting dilution assays were utilized to corroborate the in vitro findings. BFT-pretreated MCF7 cells exhibit increased tumor growth and form multifocal tumors in mice. MCF10A-KRas cells, pretreated with BFT, also showed increased tumor progression and multifocal tumors in mice. In vivo limiting dilution assay using breast tumors from BFT-pretreated MCF7 cells exhibited a striking increase in tumor-initiating cells. Follow-up analyses of these tumors demonstrated increased migratory, invasive, and mammospheres-forming behavior, confirming that brief BFT exposure elicits long-term molecular changes. Collectively, these findings present the first in vitro and in vivo evidence to show that Bacteroides fragilis toxin induces EMT, invasion/migration and stem cell-like phenotype and leads to concomitant activation of Notch and β-catenin axes. This abstract is also being presented as Poster A66. Citation Format: Sheetal Parida, Shaoguang Wu, Nethaji Muniraj, Sumit Siddharth, Arumugam Nagaligam, Cynthia L. Sears, Dipali Sharma. Bacteroides fragilis toxin induces epithelial-to-mesenchymal transition and stem-like phenotype in breast epithelial cells and concomitantly activates Notch1 and βcatenin axes [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr PR02.

Qualitative Evaluation of Virtual Touch Imaging Quantification: A Simple and Useful Method in the Diagnosis of Breast Lesions.

ObjectiveTo test the value of qualitative virtual touch imaging quantification (VTIQ) features in differentiating benign from malignant breast lesions.MethodsFrom November 2016 to August 2017, 230 lesions were subjected to conventional US and virtual touch imaging quantification before biopsy. The maximum shear wave velocity (SWVmax) was measured using a standardized method. Qualitative VTIQ features, including the “stiff rim” sign and color pattern classification, were assessed according to a binary classification. The sensitivity, specificity and area under the receiver operating curve (AUC) of Breast Imaging Reporting and Data System (BI-RADS), SWVmax, qualitative VTIQ features, and combined data were compared.ResultsAmong the 230 breast lesions, 150 were benign and 80 were malignant. Compared to the benign lesions, the malignant ones had higher SWVmax values and were more likely to show the “stiff rim” sign and VTIQ pattern 2 (P <0.001 for all). The AUC value was 0.885 for the qualitative VTIQ combination (the presence of the “stiff rim” sign and/or the display of VTIQ pattern 2), similar to that for SWVmax (P=0.472). BI-RADS combined with the qualitative VTIQ combination and with SWVmax yielded similar results, including significantly higher AUC values (P = 0.018 and 0.014, respectively), significantly higher specificities (P<0.001 for both), and nonsignificantly decreased sensitivities (P = 0.249 for both) compared to BI-RADS alone.ConclusionThe dual-category classification of qualitative VTIQ features according to the presence of the “stiff rim” sign and/or the classification of VTIQ pattern 2 is a simple and useful method that may be representative of quantitative VTIQ parameters in the evaluation of breast masses.

Risk Factors for Infectious Complications after Surgical Treatment of Spinal Metastases in Patients with Breast and Kidney Cancer

Relevance. In patients with the most common malignant new growth such as breast, kidney and lung cancer the rate of spinal metastases amounts to 70%. Increasing number of surgical procedures results in growing frequency of postoperative complications including surgical site infection (SSI) which do not only deteriorate the quality of patient’s life but change the timelines for renewal of therapy for the primary disease. Study design — case control study. Purpose of the study — to identify key risk factors as well as impact of tumor therapy on development of infectious complications in patients with breast and kidney cancer after surgical management of metastatic spine lesion. Materials and Methods. The authors collected and compiled the data on 2023 oncological patients who underwent specialized neuroorthopaedic treatment in the period from 2000 until 2017 due to tumor spine lesions. Inclusion criteria: malignant breast and kidney tumors with spine metastases, continued systemic tumor therapy, decompression and stabilization spine surgery irrespective of used implants. Patients that corresponded to inclusion criteria were divided into two groups. The first (main) group included patients (n = 22) with infectious complications after surgery. The second (control) group (n = 23) was formed by propensity score matching. Results. The analysis of obtained data resulted in identification of severe significant factors (р0.05): diabetes mellitus, postoperative liquorrhea, certain classes by ASA (3, 4) and ECOG (2, 3, 4) scales, volume of blood loss, time of surgery and type of tumor therapy. However, the three latter have the highest statistical significance (р0.01): surgical factors (blood loss volume and time of procedure) and type of tumor therapy. Conclusion. Postoperative SSI remains a common severe complication after surgeries due to metastatic spine lesions. The causes of postoperative infection after tumor resection are compromised immune status of the patient; long time of procedure with heavy blood loss and adjuvant tumor therapy.

Automatic classification of breast lesions usingTransfer Learning

Breast lesions are one of the most common types of lesions among women in Brazil and worldwide, accounting forabout 28% of new cases each year. These lesions may have Benignor Malignant behaviors. In this work, a computational method-ology for image classification was developed to differentiate malignant and benign lesions breast, aiming at low computational cost and good efficiency. In our approach, different Convolutional Neural Networks architectures and several classifiers were tested. Transfer Learning was employed to deal with the limitation of the small number of images in the database, reaching an accuracy of 81.73%, a sensitivity of 85.66%, a specificity of 78.40%, Kappa of 0.63 and ROC curve of 0.82. Finally, it is believed that theproposed methodology can integrate a CAD tool acting as patient screening or providing a second opinion to the specialist.

Breast tumour size as a predictor of hemostatic system status and endothelial function in dogs

The high level of morbidity of dogs from mammary gland tumours and deficiencies in the pathogenesis give relevance to study of the disorders of the hemostatic system for predicting the neoplasia process. Our research concerned the determination of markers of the hemostatic system and endothelial function at different sizes (≤ 2, 5–7 and &gt;10 cm) of benign (n = 28) and malignant (n = 27) breast tumours in dogs, taking into account their histological structure. The progression of the tumour process was accompanied by a 1.24–1.81 times increase in the fibrinogen content in the benign forms and 1.39–2.38 times in malignant against the background of progressive excess accumulation of soluble fibrin. The results of coagulation tests indicate that the increase in blood coagulation, which correlates with the magnitude of tumours, occurred mainly externally. In malignant neoplasms reliable, compared to clinically healthy dogs, reduction of total fibrinolytic activity occurred due to deficient inhibition of tissue plasminogen activator by 1.62–2.03 times. Increase in the size of benign neoplasms was accompanied by increased activity of the α1-inhibitor of proteinases, and in malignant – only in tumours of the size of 5–7 cm, against the background of its decrease in dogs with small and giant neoplasia. The progression of the disease was characterized by an increase in the content of α2-macroglobulin in benign lesions from 1.19–1.24 times, malignant – from 1.25–2.03 times. At the same time, there was a deepening of oxidative stress, as evidenced by the excess accumulation in the blood of malondialdehyde by 1.29–1.70 times in benign tumours, and 1.58–2.73 times in malignant. Regardless of the pathomorphologic form, the magnitude of the neoplasia foci is directly correlated with excess accumulation of nitric oxide and the level of hypoproteinemia, and vice versa – the content of ceruloplasmin. Further research should be undertaken to study the hemostatic status in certain nosological forms of breast tumours in dogs, which will improve the diagnosis and development of effective treatment protocols.

Comparative Analysis of the Quantitative Parameter Method and Elasticity Color Mode Method for Real-Time Shear Wave Elastography in the Diagnosis of Benign and Malignant Solid Breast Lesions

Ultrasound elastography is a new technique that has been developed rapidly in recent years and has attracted much attention. It can reveal the histological characteristics of a lesion by detecting the elasticity of the tissue

Abstract 2834: Bacteroides fragilis: A potential pathogen orchestrating EMT and stemness in breast epithelial cells via concomitant activation of Notch and βcatenin axes

Abstract Background: Last decade established significant contributions of microbiome to organ specific cancers. A few recent studies suggested the existence of distinct breast microbiota and a shift in microbial community composition in diseased breast compared to normal breast. However, their functional impact and underlying mechanisms are unknown. Present study was designed to examine the contribution of pro-carcinogenic bacteria in breast cancer initiation, growth and progression. Results: Utilizing extensive data mining and metagenomic analyses, we discovered presence of toxin producing Bacteriodes fragilis in malignant breast. B. fragilis is known for its potential to initiate and/or promote colon cancer. Its pathogenicity has been attributed to its unique toxin BFT. Mice infected with B. fragilis exhibited significant circulating BFT and distinct morphological alterations in mammary gland. While no changes were observed in cell growth and clonogenicity upon BFT treatment, significant increase in migration and invasion potential and decreased adhesion of MCF10A and MCF7 cells were observed. BFT-treated cells displayed acquisition of fibroblast-like appearance and increased formation of pseudopodia/microtentacles emanating from the cell membrane along with molecular markers of epithelial-to-mesenchymal transition. Decreased expression of epithelial marker, E-cadherin along with elevated levels of mesenchymal markers, N-cadherin and vimentin were observed. BFT also increased expression of EMT-related transcription factors, Snail, Slug and Twist. BFT-treated cells attained stem cell-like phenotype exhibiting an increased ability to form secondary and tertiary mammospheres and elevated expression of pluripotency-factors (Oct4, Nanog and Sox2). Mechanistic studies showed BFT induced expression and nuclear translocation of cleaved NOTCH1 and βcatenin resulting in activation of downstream targets. Inhibition of Notch1 and βcatenin using γ-secretase inhibitor and ICG001 successfully inhibited functional effects of BFT. Further, BFT-pretreated MCF7 cells exhibit increased tumor growth and form multifocal tumors in mice. MCF10A-KRas cells, pretreated with BFT, also showed increased tumor progression and multifocal tumors in mice. In vivo limiting dilution assay using breast tumors from BFT-pretreated MCF7 cells exhibited a striking increase in tumor-initiating cells. Follow-up analyses of these tumors demonstrated increased migratory, invasive, and mammospheres-forming behavior confirming that brief BFT exposure elicits long-term molecular changes. Conclusion: Collectively, these findings present the first in vitro and in vivo evidence to show that Bacteriodes fragilis Toxin induces EMT, invasion/migration and stem cell-like phenotype and leads to concomitant activation of Notch and βcatenin axes. Citation Format: Sheetal Parida, Shaoguang Wu, Nethaji Muniraj, Sumit Siddharth, Arumugam Nagaligam, Christina Hum, Panagiotis Mistriotis, Konstantinos Konstantopoulos, Cynthia L. Sears, Dipali Sharma. Bacteroides fragilis: A potential pathogen orchestrating EMT and stemness in breast epithelial cells via concomitant activation of Notch and βcatenin axes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2834.

Use of single-photon emission computed tomography combined with computed tomography for breast cancer diagnosis

Objective. To evaluate the efficacy of single-photon emission computed tomography combined with computed tomography (SPECT-CT) in the diagnosis of breast cancer (BC).Materials and methods. We performed 44 SPECT-CT examinations in 40 women with malignant breast neoplasms.Study Results. The obtained data enabled us to calculate the characteristics of SPECT-CT in the evaluation of the primary tumor. The sensitivity, specificity, and diagnostic accuracy of the method were 88.1 %, 78.3 %, and 88.0 %, respectively. When the method was used to detect nodal and distant metastases, these characteristics were 87.3 %, 76.8 %, 86.9 % and 82.6 %, 73.1 %, 81.4 %, respectively.Conclusion. SPECT-CT is one of the most accurate methods to assess the primary tumor, lymph node involvement and distant metastases in BC.

Comparative analysis of confocal microscopy on fresh breast core needle biopsies and conventional histology

BackgroundEvaluation of core needle biopsies (CNB) is a standard procedure for the diagnosis of breast cancer. However, tissue processing and image preparation is a time- consuming procedure and instant on-site availability of high-quality images could substantially improve the efficacy of the diagnostic procedure. Conventional microscopic methods, such as frozen section analysis (FSA) for detection of malignant cells still have clear disadvantages. In the present study, we tested a confocal microscopy scanner on fresh tissue from CNB with intention to develop an alternative device to FSA in clinical practice.Patients and methodsIn 24 patients with suspicious breast lesions standard of care image-guided biopsies were performed. Confocal images have been obtained using the Histolog™ Scanner and evaluated by two independent pathologists. Hematoxylin-Eosin (H&E) histological sections of the biopsies were routinely processed in a blinded fashion with respect to the confocal images.ResultsIn total 42 confocal images were generated from 24 biopsy specimens, and available for analysis within a few minutes of taking the biopsy. This resulted in 2 × 42 = 84 pathologic evaluations. In four cases, a pathologic diagnosis was not possible with confocal microscopy. An exact correlation based on the B-classification was reached in 41 out of 80 examinations and in another 35 cases in a broader sense of correspondence definition (i.e. malignant vs. benign).ConclusionsAs a reliable on-site method, the Histolog™ Scanner provides a visualization of cellular details equivalent to the H&E standards, permitting rapid and accurate diagnosis of malignant and benign breast lesions. Furthermore, this device offers great potential for immediate margin analysis of specimen in breast conserving therapy.

Diffusion-Weighted MRI of Breast Cancer: Improved Lesion Visibility and Image Quality Using Synthetic b-Values.

BackgroundDiffusion‐weighted imaging (DWI) is an MRI technique with the potential to serve as an unenhanced breast cancer detection tool. Synthetic b‐values produce images with high diffusion weighting to suppress residual background signal, while avoiding additional measurement times and reducing artifacts.PurposeTo compare acquired DWI images (at b = 850 s/mm2) and different synthetic b‐values (at b = 1000–2000 s/mm2) in terms of lesion visibility, image quality, and tumor‐to‐tissue contrast in patients with malignant breast tumors.Study TypeRetrospective.PopulationFifty‐three females with malignant breast lesions.Field Strength/SequenceT2w, DWI EPI with STIR fat‐suppression, and dynamic contrast‐enhanced T1w at 3T.AssessmentFrom acquired images using b‐values of 50 and 850 s/mm2, synthetic images were calculated at b = 1000, 1200, 1400, 1600, 1800, and 2000 s/mm2. Four readers independently rated image quality, lesion visibility, preferred b‐value, as well as the lowest and highest b‐value, over the range of b‐values tested, to provide a diagnostic image.Statistical TestsMedians and mean ranks were calculated and compared using the Friedman test and Wilcoxon signed‐rank test. Reproducibility was analyzed by intraclass correlation (ICC), Fleiss, and Cohen's κ.ResultsRelative signal‐to‐noise and contrast‐to‐noise ratios decreased with increasing b‐values, while the signal‐intensity ratio between tumor and tissue increased significantly (P < 0.001). Intermediate b‐values (1200–1800 s/mm2) were rated best concerning image quality and lesion visibility; the preferred b‐value mostly lay at 1200–1600 s/mm2. Lowest and highest acceptable b‐values were 850 s/mm2 and 2000 s/mm2. Interreader agreement was moderate to high concerning image quality (ICC: 0.50–0.67) and lesion visibility (0.70–0.93), but poor concerning preferred and acceptable b‐values (κ = 0.032–0.446).Data ConclusionSynthetically increased b‐values may be a way to improve tumor‐to‐tissue contrast, lesion visibility, and image quality of breast DWI, while avoiding the disadvantages of performing DWI at very high b‐values. Level of Evidence: 3 Technical Efficacy: Stage 2J. Magn. Reson. Imaging 2019;50:1754–1761.

Multiplex PCR analysis of apocrine lesions shows frequent PI3K-AKT pathway mutations in both benign and malignant apocrine breast tumors.

Pathological diagnosis of intraductal apocrine lesions can be challenging, because even benign apocrine lesions often show atypical cytology, and immunohistochemistry is of little assistance. A new diagnostic method for apocrine lesions is desirable. The mutations present in apocrine lesions have not been well studied. We performed a MassARRAY multiplex polymerase chain reaction (PCR) study of benign and malignant apocrine lesions, which included 152 mutations of 18 genes. We found that four of 11 benign lesions showed AKT1 or PIK3CA mutations, one of four noninvasive apocrine carcinomas showed a FBX4 mutation, two of 15 invasive apocrine carcinomas showed a PIK3CA mutation, and one invasive apocrine carcinoma showed both PIK3CA and TP53 mutations. The mutation frequency did not differ significantly between benign and malignant lesions (p = 0.683). We demonstrated that both benign and malignant apocrine lesions may contain mutations of genes in the PI3K-AKT pathway, this pathway could be a good therapeutic target of these diseases.

Practical exploration and research of Watson for oncology clinical decision support system in real-world and localized practice.

e18304 Background: Watson for Oncology (WFO) is a cognitive computing system developed by the IBM and trained by Memorial Sloan Kettering Cancer Center to provide evidence-based treatment recommendation for cancer patients. This study is to explore the impact on clinical treatment decision and patient communication based on the standard workflow we established of applying WFO in real-world clinical practice. Methods: From May 2018 to January 2019, 57 patients with advanced malignant tumors, including lung, breast, colorectal, gastric, esophageal, liver, ovarian and endometrial cancer, treated at Oncology Department of Beijing Chaoyang Integrative Medicine Emergency Medical Center were included in the study, of which 28 males and 29 females aged 29-89 years old. Treatment plan of each patient generated by WFO was compared with NCCN guidelines and Chinese local CSCO guidelines, and then evaluated and determined by MDT for final decision. Proportion of WFO recommendation fully or partially followed by clinicians was analyzed. Moreover, survey by questionnaire was conducted to collect feedback from these patients on their satisfaction of treatment and communication process. Results: Of all 57 patients enrolled, WFO’s options were highly concordant with NCCN guidelines and CSCO guidelines, and the finally adopted treatment plans after deliberately evaluated and discussed by MDT were 100% in the WFO recommended or for consideration category. Patient satisfaction and the compliance with instruction was significantly improved compared with historical data. Localization was most required in gastric, liver and esophageal cancer due to local drugs and therapies not available in WFO. Conclusions: This study verified that the artificial intelligent clinical decision supporting system can be applied in Chinese clinical practice to achieve high patient satisfaction and good efficacy. WFO meets Chinese and international standards for treatment, which will promote the standardized development of cancer treatment in China. Optimization, iteration, upgrading and localization are required for better practicability of WFO.

Experience and impacts of remote telepathology consultation in improving cancer patients’ diagnosis and management in China.

e18337 Background: Over the past 4 years, we have conducted over 30 telepathology consultation cases from China covering a wide spectrum of cancers. Through our experience, we realized that the diagnostic accuracy and report integrity from Chinese pathologists is lower than US standards. which brings great concern about diagnosis, treatment and prognosis of cancer patients in China. Methods: Consultation cases were submitted by Chinese patients, their family members or physicians. Specifically, all clinical reports were sent for consultation including imaging studies, laboratory tests, pathology reports, and histology slides for review which covered a wide spectrum of malignant neoplasms, including lung, esophagus, stomach, colon, ovary, breast, prostate, neuroendocrine, melanoma, head/neck, and soft tissue sarcoma; for most cases (27/30) additional immunostaining or molecular tests were performed. All cases and reports were analyzed at top pathology institutions in the United States, including Memorial Sloan Kettering Cancer Center, Johns Hopkins Hospital, or Brigham Women’s Hospital/Beth Israel Medical Center. Following the review of the original materials, final diagnostic reports between the Chinese and US medical centers were compared for major discrepancies in diagnosis. Results: All consulted cases had major discrepancies in diagnosis (100%). Specifically, 75% of cases had improper cancer classifications or grading, 2 cases had improper quality control in immunostains rendering interpretation questionable; 3 cases lacked appropriate molecular tests; finally, 29/30 (97%) of cases lacked cancer staging based on CAP/AJCC/UICC TNM classification of cancer report protocols. Conclusions: Our consultation experiences demonstrated great discrepancies in malignant neoplasm diagnosis and reports among pathologists between the US and China. These findings raise concerns for Chinese patient’s clinical management and prognoses; moreover, they might help explain the observed difference in cancer patient's total 5-year survival between China and USA (40% vs 70%). These findings emphasize the importance of providing support and consultations for pathologists in China, especially those who practice in non-metropolitan, small county or rural local hospitals which lack updated information, technical and academic supports.

All Benign and Malignant Apocrine Breast Lesions Over-Express Claudin 1 and 3 and Are Negative for Claudin 4

Invasive apocrine carcinoma of the breast is an uncommon triple negative tumour that lacks a specific therapeutic target. Apocrine metaplasia of the breast shares common morphological features with apocrine carcinoma, and was previously found to consistently over-express claudin 1 and to lack claudin 4. This study was aimed at finding whether apocrine carcinoma, and other related apocrine breast lesions, have similar claudin profile. The immunohistochemical expression of claudin 1, 3 and 4 was studied in 11 cases of in situ and invasive apocrine breast carcinoma, 7 benign apocrine lesions and 45 consecutive morphologically non-apocrine triple negative breast carcinomas. All cases were also immunostained for Gross Cystic Disease Fluid Protein-15 (GCDFP-15), a marker for apocrine differentiation. Apocrine breast lesions maintained their expression pattern from benign through DCIS to invasive carcinoma; all showing strong expression of claudin 1 and 3 and absence of claudin 4. The same pattern of expression was seen in 2 out of the 45 morphologically non-apocrine tumours, but both showed strong positive staining for GCDFP-15. It is concluded that all benign and malignant apocrine lesions of the breast have a consistent pattern of claudin 1, 3 and 4 expression, suggesting the presence of a specific pathway for the development of invasive apocrine carcinoma. The over-expression of claudin 1 and 3 may have therapeutic implications as targets for managing apocrine cancers.

Survey of dentin sialophosphoprotein and its cognate matrix metalloproteinase-20 in human cancers.

BackgroundMatrix metalloproteinases‐20 (MMP20) expression is widely regarded as tooth specific, with expression limited to dental hard tissues. Recently, we reported MMP20 expression and interaction with dentin sialophosphoprotein (DSPP), a member of the Small Integrin Binding Ligand N‐linked Glycoproteins (SIBLINGs), in human oral squamous cell carcinoma (OSCC) and dysplastic oral premalignant lesions (OPLs), suggesting a role for MMP20‐DSPP interaction in oral carcinogenesis.MethodsThis study aimed to survey the expression of MMP20 and its cognate DSPP partner in the breast, colon, prostate, thyroid, and cervical neoplasms. Using commercially available tissue microarrays (TMAs) and cell lines, we performed immunohistochemistry, immunofluorescence, proximity ligation assay, and western blot experiments to determine the expressions of MMP20 and DSPP in the breast, colon, prostate, thyroid, cervical neoplasms, and their normal counterparts.ResultsSignificantly high expression levels of MMP20 and DSPP were observed in the malignant breast, colon, prostate, thyroid, and cervical neoplasms compared with their benign and normal counterparts. Furthermore, MMP20 levels increased with advanced stages of colon and thyroid cancers. DSPP expression increased significantly with tumor stage in all cancers examined.ConclusionsThe co‐localization and potential MMP20‐DSPP interaction previously reported in oral cancers are present in other cancers. These results suggest MMP20‐DSPP pairing as a potential marker of disease activity in some epithelial cancers with diagnostic and prognostic implications.

Methylation Changes of Primary Tumors, Monolayer, and Spheroid Tissue Culture Environments in Malignant Melanoma and Breast Carcinoma.

Epigenetic changes have major role in the normal development and programming of gene expression. Aberrant methylation results in carcinogenesis. The primary objective of our study is to determine whether primary tumor tissue and cultured tumor cells in 2D and 3D tissue culture systems have the same methylation signature for PAX5, TMPRSS2, and SBDS. These findings will play an important role in developing in vitro model system to understand the effect of methylation inhibitors on primary tumor tissue. In a previous study PAX5, TMPRSS2, and SBDS genes that we are investigating were reported to be methylated more than 60% in breast cancer and malignant melanoma cell lines. However, these genes have never been studied in primary tumor tissues. Thus, primary tumor tissues of breast cancer and malignant melanoma were first grown in 2D and 3D cultures. Then these two types of tumor tissues and their 2D and 3D cultures were investigated for changes considering methylation levels in PAX5, TMPRSS2, and SBDS genes using real-time polymerase chain reaction. No differences were observed in the primary tissues and culture systems for both PAX5 and TMPRSS2 in malignant melanoma tissues. We found that PAX5 gene was an efficient marker to measure the effects of methylation inhibitors for in vitro systems for malignant melanoma tissue.

Diagnostic Accuracy of Shear Wave Elastography in Differentiation Between Benign and Malignant Solid Breast Masses Compared with Strain Elastography

The aim of this study was to evaluate the diagnostic performance of shear wave elastography by acoustic radiation force impulse (ARFI) elastography in differentiating malignant and benign breast Lesions in comparison with strain elastography and B mode ultrasound. This was prospective study, we used the commercially available eSie touch elastography imaging. In the shear wave elastography (SWE) we had two modes, the virtual touch imaging (VTI) with interpretation with the 5 points elasticity score and virtual touch quantification (VTQ) technique with the calculation of the Shear wave velocity (SWV). The study included 142 solid breast masses, of them 75 (52.8%) were benign and 67 (47.2%) were malignant. The mean shear wave velocity differed significantly between the benign and malignant groups (2.4+1.3m/sec and 7.3+2.2m/sec respectively, P value <0.0001). the sensitivity, specificity, PPV, NPV and accuracy of strain (eSie touch) elastography score was 83.1%, 88.73%, 88.06%, 84% and 85.92% respectively, which was less than the elastography score by ARFI (92.42%, 92.11%, 91.04%, 93.33% and 92.25% respectively) and less than the VTQ (SWV) which was 94.03%, 95.95%, 95.45% and 95.04% respectively. We concluded that Both the SWE and SE elastography showed significant difference between the benign and malignant masses, and both has added value above B mode ultrasound during routine examination. Shear wave elastography had higher sensitivity and specificity than SE, and less false negative and false positive results. The quantitative SWE (SWV) had the highest diagnostic performance among the different studied techniques.

Biochemical Investigation of Tissue Oxidative Stress and Angiogenesis with Associated Trace Elements in Breast Disease Patients in Uttarakhand, India.

Breast cancer is a heterogeneous cancer with diverse clinical symptoms and an ambiguous molecular spectrum. Oxidative damage, antioxidant activity, and angiogenesis combine to form significant complex factors that stimulate breast cancer development and progression. This study is designed to determine level changes in total antioxidant status and markers of lipid peroxidation melondialdehyde (MDA) and angiogenesis vascular endothelial growth factor (VEGF) along with related micronutrients of copper, zinc, magnesium, and iron in malignant and benign breast disease tissue extracts. We assess specificity and sensitivity of those markers using the area under the curve of receiver operator characteristic (ROC) curve analysis. Association studies are done with correlation analysis. The tissue extract level of MDA markers is found to be significantly higher (14.118 ± 1.47 nmol/g tissue; p < 0.05), with significantly depleted levels of antioxidants (5.983 ± 1.661 nmol/g tissue; p < 0.05). The tissue VEGF level also significantly increases in a diseased condition (512.466 ± 5.661 pg/mg tissue) versus the nondiseased condition (422.433 ± 13.615 pg/mg tissue). Related trace-element levels show a significant mixed pattern among studied groups. VEGF emerges as the best discriminatory biomarker of breast cancer presence, in accordance with ROC analysis. Oxidative stress and angiogenesis are found to be important factors in breast cancer development. This study forms the basis for future studies that focus on the relationship between roles of indices studied and cancer induction.

Nodal induces epithelial-mesenchymal transition in gallbladder carcinoma via the Smad2/4 pathway

Objective To study the expressions of Nodal in normal gallbladder, and gallbladders with cholelithiasis, cholecystitis and carcinoma; and to study the impact of inhibiting or promoting Nodal expressions in gallbladder carcinoma on the Smad2/4 pathway and epithelial-mesenchymal transition. Methods Immunohistochemistry was used to detect the expressions and distributions of Nodal protein in 30 normal gallbladders, 96 simple cholecystitis/calculous cholecystitis specimens and 42 gallbladder carcinoma specimens. The mRNA and protein expressions of Nodal in normal and malignant gallbladder mucosal epithelium cells and breast cancer cells were detected by RT-PCR, western blotting and wound healing tests. The impact of activating agents and inhibitors on the expression levels of Nodal and its signaling pathway Smad2/4 and EMT-related proteins were analyzed. Results Immunohistochemistry showed that the positive rates of Nodal in the gallbladder cancer group was significantly higher than that in the gallbladder stone group and the normal gallbladder group. The results were 83.3% (35/42), 44.8% (43/96), 6.7% (2/30) respectively (P<0.01). RT-PCR and Western blotting showed the expressions of Nodal in gallbladder carcinoma cells were higher than normal gallbladder cells (P<0.05). After using rhNodal to up regulate the Nodal expression, the Smad2 protein phosphorylation was promoted and the EMT associated proteins were up-regulated. After using the inhibitor SB431542 to suppress the Nodal expression, the Smad2 protein phosphorylation decreased and the EMT associated proteins were down-regulated. Conclusions The expression of Nodal was closely related to cell proliferation and metastasis in gallbladder carcinoma. Tumor progression was promoted via the smad2/4 pathway through epithelial-mesenchymal transition. Key words: Gallbladder carcinoma, Gallstone; Transforming growth factor-beta, Nodal; Smad2/4, signal conduction; Epithelial-mesenchymal transition