Non-invasive Breast Carcinoma Research Articles

Modern visualization diagnostic methods of non-invasive breast carcinomas (review of literature)

Relevance. Breast carcinomas (BC) remain one of the most actual problems of modern oncology. According to statistics, the incidence of BC is steadily increasing, making it the most common cancer pathology among women. In this situation, the aspect of diagnosing BC at early, non-invasive stages, is certainly important, which still reduces mortality, increases the possibility of organ-preserving treatment, duration and quality of life of patients.The purpose of study. The purpose of study is to investigate and perform a comparative analysis of imaging techniques for the diagnosis of non-invasive breast carcinomas.Materials and methods. Modern Russian and foreign literature about the diagnosis of non-invasive breast carcinomas was analyzed. Publications not older than 7 years published in specialized medical editions were taken into account Results. Not only main, but also new, promising imaging modalities that are not currently part of routine practice were analyzed. The main imaging patterns in non-invasive breast carcinomas (DCIS, LCIS and Paget's breast cancer), feasibility and prognostic value of certain diagnostic methods in different nosologies of this disease were also discussed.Conclusion. The statistics provided explain the importance of breast carcinomas problem, as well as the relevance of its diagnosis at non-invasive stages. According to the authors, the issue of accurate diagnosis of breast cancer in situ is subject to further discussion and study, but we would like to note that when non-invasive breast carcinoma is suspected and doubtful imaging results are obtained, one should not limit oneself only to routine methods of imaging studies, but expand further diagnostic tactics until accurate results are obtained and a final diagnosis is made.

Outcomes in patients with non-invasive breast carcinoma.

Non-invasive breast carcinoma is considered to be localized disease and is distinguished from invasive ductal and lobular carcinomas. The local recurrence of non-invasive carcinoma after surgery may lead to development of invasive carcinoma and promote distant metastasis, which worsens the prognosis for breast cancer mortality. The distant metastasis of non-invasive carcinoma may involve the ductal microvasculature without invasion. The outcomes of non-invasive breast carcinoma were examined in this retrospective cohort study. Of 872 primary breast cancers diagnosed at a single center between May 2008 and March 2022, 93 (10.6%) were found to be non-invasive carcinomas and were examined in this study. The breast cancer recurrence and survival rates of patients with non-invasive carcinoma were analyzed retrospectively. The median follow-up period was 1891 (range, 5-4804) days. All patients underwent surgical treatment [mastectomy with sentinel lymph node biopsy (SLNB) and partial mastectomy with or without SLNB, tumorectomy, and microdochectomy]. Postoperatively, radiation therapy was administered to 73 (78.4%) of the patients and endocrine therapy was administered to 64 (81.0%) of 79 patients with hormone-receptor positivity. Of 26 patients who underwent partial mastectomy with SLNB, 24 (92.3%) showed isolated tumor cells in the SLNs on one-step nucleic acid amplification. Local recurrence was observed in three (0.3%) patients; no distant metastasis was observed. One patient died of a noncancerous disease. The overall survival rate was 98.0% and the breast cancer-specific survival rate was 100.0%. Non-invasive breast carcinoma, like invasive breast carcinoma, causes local recurrence, but has a good prognosis without distant metastasis. The clinical significance of isolated tumor cells in the SLNs as a systemic component of non-invasive breast carcinoma remains to be elucidated.

Circulatory RNA based non-invasive detection and diagnosis of breast cancer

Breast carcinoma is the most prevalent and lethal form of carcinoma in women, as well as the most prominent source of cancer-related mortality in women. The most widespread form of non-invasive breast carcinoma is ductal carcinoma in situ (DCIS). Early detection of breast carcinoma improves the chances of rehabilitation and survival, lowering mortality rates. Limitations of mammography as a screening test have made it imperative to develop new, non-invasive, and more convenient alternatives for the prognosis of breast carcinoma. Circulatory RNAs (miRNAs) have been included in a novel category of non-coding, endogenous, small RNA fragments that can be considered gene expression regulators, influencing key processes in cancer development. Peculiar expressions and firmness of circulatory RNAs exhibit a promising possibility of their use as biomarkers in the prognosis of mammary tumor.

Quantitative classification of invasive and noninvasive breast cancer using dynamic magnetic resonance imaging of the mammary gland.

ObjectivesBreast cancers are classified as invasive or noninvasive based on histopathological findings. Although time-intensity curve (TIC) analysis using magnetic resonance imaging (MRI) can differentiate benign from malignant disease, its diagnostic ability to quantitatively distinguish between invasive and noninvasive breast cancers has not been determined. In this study, we evaluated the ability of TIC analysis of dynamic MRI data (MRI-TIC) to distinguish between invasive and noninvasive breast cancers.Material and MethodsWe collected and analyzed data for 429 cases of epithelial invasive and noninvasive breast carcinomas. TIC features were extracted in washout areas suggestive of malignancy.ResultsThe graph determining the positive diagnosis rate for invasive and noninvasive cases revealed that the cut-off θi/ni value was 21.6° (invasive: θw > 21.6°, noninvasive: θw ≤ 21.6°). Tissues were classified as invasive or noninvasive using this cut-off value, and each result was compared with the histopathological diagnosis. Using this method, the accuracy of tissue classification by MRI-TIC was 88.6% (380/429), which was higher than that using ultrasound (73.4%, 315/429).ConclusionMRI-TIC is effective for the classification of invasive vs. noninvasive breast cancer.

Artesunate-induced Cellular Effects Are Mediated by Specific EPH Receptors and Ephrin Ligands in Breast Carcinoma Cells

The aberrant regulation of erythropoietin-producing hepatocellular carcinoma (EPH) receptors and ephrin ligands has been implicated in breast carcinoma, and artesunate has been shown to have anticancer effects. The aim of this study was to characterize the involvement of EPH receptors and ephrin ligands in mediating artesunate (ART)-induced growth suppression of normal breast cells and breast carcinoma cell lines. The normal breast epithelial cells (MCF10A), non-invasive ductal breast carcinoma cells (MCF7), and invasive triple-negative breast carcinoma cells (MDA-MB-231) were grown in the absence or the presence of different concentrations of artesunate. The cells were counted, and total RNA was isolated. The abundance of transcripts corresponding to EPH receptors and ephrin ligands was determined by quantitative polymerase chain reaction. Cell viability was significantly reduced when cells were treated with artesunate, with MDA-MB-231 cells having the highest sensitivity. Artesunate had no significant effect on transcription of EPH/ephrins in MCF10A cells, but markedly increased EPHA8, EPHA10, EPHB6 and ephrin-A2 expression in MCF7 cells, and significantly increased EPHA3 and EPHA10 expression while reducing that of EPHA7 and ephrin-A3 in MDA-MB-231 cells. The relative changes in artesunate-treated MCF7 and MDA-MB-231 cells as compared to similarly treated MCF10A cells allow us to implicate combinatorial expression and receptor interactions for EPH receptor-mediated signal transduction that converges into pathways responsible for cell growth, proliferation, and apoptosis. Specifically, the alterations in EPHA7, EPHA8, EPHA10 and EPHB6 transcripts appear to be important participants in artesunate-mediated cellular effects.

Abstract 5949: Artesunate influences the transcripts for Eph receptors and ephrin ligands in breast cancer cells

Abstract Purpose: Artesunate (ART), an anti-malarial drug, has been shown to induce apoptosis in breast cancer cells. The current study was conducted to explore the involvement of Eph receptor and ephrin ligand transcript levels in mediating apoptotic functions of ART in three phenotypically distinct breast cell lines. The specific involvement Eph receptors and ephrin ligands in apoptosis may potentially lead to novel approaches for therapeutic intervention of breast cancer. Methods: We cultured three cell lines that represent non-tumorigenic breast epithelium (MCF10A), non-invasive breast carcinoma (MCF7), and invasive triple negative breast carcinoma (MDA- MB-231) in the presence of varying concentrations of ART. Total RNA was isolated from untreated and ART-treated cell lines, and the relative abundance of Eph receptor and ephrin ligand transcripts was determined by qPCR. The changes in selected transcripts were confirmed by immunocytochemistry and western blot analysis. Results: Our study revealed that the transcripts corresponding to various Eph receptors and ephrin ligands are differentially expressed in a cell line specific manner in ART-treated cells. Specifically, EphA5 was upregulated and EphB4 was significantly downregulated in MCF10A cells. However, MDA-MB-231 cells were distinguished by upregulation of ephrinA2 and downregulation of EphA7, respectively. MCF7 also showed distinct response to ART treatment. Combinatorial alterations in transcript abundance for a subset of Eph receptors and ephrin ligands are likely to provide meaningful insights in the apoptotic effects of ART in breast cancer cells. Conclusions: A variety of experiments directly or indirectly support the role of Eph receptors and ephrin ligands in apoptosis of numerous cell types. To our knowledge, there have been no conclusive studies to implicate these molecules in ART-mediated cell death. We have demonstrated cell line-specific alterations in the levels of Eph/ephrin transcripts in cells primed for apoptosis by ART-treatment. Specifically, EphA5, EphA7, EphB4 and ephrin A2 appear to transduce apoptotic signals in ART-treated cells. These observations set the basis for pursuing additional mechanistic investigations to decipher novel mediators of apoptosis in breast cancer cells. Citation Format: Tanin Zadeh, Mariana Lucero, Raj Kandpal. Artesunate influences the transcripts for Eph receptors and ephrin ligands in breast cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5949.

Abstract 5107: DDR1 and MT1-MMP expression levels are major determinants of 3D type I collagen-induced apoptosis in basal-like breast carcinoma

Abstract Type I collagen is the major adhesive component in breast interstitial stroma, which represents the first barrier against tumor cell invasion after basement-membrane degradation. Among cellular receptors, type I collagen is able to activate discoidin domain receptors DDR1 and DDR2. We have previously shown that in 3D collagen matrix, DDR1 plays a key role as it promotes cell growth suppression and apoptosis through the upregulation of the pro-apoptotic mediator BIK in noninvasive luminal-like breast carcinoma cells. We have also shown that MT1-MMP is able to rescue these cells and protect them against the effects induced by collagen/DDR1/BIK axis. Our data suggested that the protective effect of MT1-MMP might be mediated through the degradation of type I collagen and/or DDR1 cleavage. Decreased DDR1 expression has been associated with the epithelial to mesenchymal transition process in breast cancer, and its overexpression in aggressive basal-like breast cancer cells reduces their invasiveness in 3D cultures and in vivo. In the present work, we propose to study the role of MT1-MMP in the resistance against collagen-induced apoptosis in basal-like breast carcinoma MDA-MB-231 cells. We aimed to investigate whether MT1-MMP depletion is able to restore apoptosis mediated by collagen/DDR1/BIK axis and to verify if such depletion is able to restore full-length DDR1 expression and phosphorylation. ShRNA strategy against MT1-MMP mRNA was able to partially restore full length DDR1 expression and phosphorylation. This was accompanied by a decrease in cell growth and an upregulation of BIK expression. This suggested that MT1-MMP expression in basal-like breast carcinoma cells, in addition to a low basal level of DDR1 expression, protects these cells against collagen-induced apoptosis via DDR1 cleavage. Since DDR1 was moderately expressed in MDA-MB-231 cells, we then investigated whether overexpression of DDR1 could be able to increase its ability to suppress cell growth and to induce apoptosis. Data showed that overexpression of DDR1 induced a decrease in cell growth and an increase in BIK expression, suggesting that moderate expression level of full length DDR1 in basal-like breast carcinoma provides them with a capacity to resist to collagen-induced cell growth suppression and apoptosis. Finally, the combined overexpression of DDR1 and depletion of MT1-MMP in MDA-MB-231 cells synergistically increased collagen-induced cell growth suppression and apoptosis to a level similar to that observed in luminal breast carcinoma. Taken together, our data suggest that during the acquisition of mesenchymal features, the low level of DDR1 expression should be considered as an important biomarker in the prognosis of basal-like breast carcinoma, conferring them a high rate of cell growth and resistance to BIK-mediated apoptosis induced by the stromal collagen. Citation Format: Charles Saby, Guillaume Collin, Maha Sinane, Emilie Buache, Laurence Van Gulick, Frédéric Saltel, Erik Maquoi, Hamid Morjani. DDR1 and MT1-MMP expression levels are major determinants of 3D type I collagen-induced apoptosis in basal-like breast carcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5107.

Abstract 4686: Stem-like cells and bulk cells in breast cell lines express distinct combinations of Eph receptors and ephrin ligands

Abstract Purpose: The maintenance of the breast cancer stem cell niche in mammary tumors has been attributed to aberrant expression of Eph receptors and ephrin ligands. The current study aims to identify Eph receptors and ephrin ligands that may contribute to the tumorigenic and invasive breast cancer phenotypes, and can potentially be used to classify tumor specific stem cells and target them for therapeutic intervention. Methods: The bulk cells and stem-like cells from MCF10A, MCF7 and MDA-MB-231 cell lines were separated based on CD44+/CD24+ and CD44+/CD24- combinations of cell surface markers, respectively. These cell lines represent non-tumorigenic breast epithelium (MCF10A), non-invasive breast carcinoma (MCF7), and invasive triple negative breast carcinoma (MDA-MB-231). The profiles of Eph receptors and ephrin ligands were determined by qPCR. The changes in selected transcripts were confirmed by immunocytochemistry and western blot analysis. Results: Our study revealed that Eph receptors and ephrin ligands have distinct expression signatures in CD44+/CD24- tumor-initiating cells relative to the bulk tumor cell population. Such combinatorial expression of these molecules in stem cells and bulk cells may modulate and maintain breast tumor phenotypes. The differential patterns of expression are evidenced by changes in qualitative as well as quantitative combinations of Eph receptors and ephrin ligands. The levels of EphA8 transcript were notably lower in CD44+/CD24- cells as compared to bulk MCF7 cells. However, EphA8 was significantly upregulated in stem cells as compared to bulk cells isolated from MDA-MB-231 cell line. Conclusion: The relative abundance of Eph receptors and ephrin ligands in stem and bulk cells provides molecular signatures of stem cells specific to tumor phenotypes. The combinatorial expression of these molecules corresponds to invasiveness of breast cancer cells, which can be potentially exploited for diagnostics and therapeutics. Citation Format: Mariana Lucero, Jaspreet Thind, Shayan Senaati, Belinda Jimenez, Tanin Zadeh, Raj P. Kandpal. Stem-like cells and bulk cells in breast cell lines express distinct combinations of Eph receptors and ephrin ligands [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 4686.

DDR1 and MT1-MMP Expression Levels Are Determinant for Triggering BIK-Mediated Apoptosis by 3D Type I Collagen Matrix in Invasive Basal-Like Breast Carcinoma Cells.

Type I collagen is the major adhesive component in breast interstitial stroma, which represents the first barrier against tumor cell invasion after basement-membrane degradation. Among cellular receptors, type I collagen is able to activate discoidin domain receptors DDR1 and DDR2. We have previously shown that in 3D collagen matrix, DDR1 plays a key role as it promotes cell growth suppression and apoptosis through the upregulation of the pro-apoptotic mediator BIK in noninvasive luminal-like breast carcinoma cells. We have also shown that MT1-MMP is able to rescue these cells and protect them against the effects induced by collagen/DDR1/BIK axis. Our data suggested that the protective effect of MT1-MMP might be mediated through the degradation of type I collagen and/or DDR1 cleavage. Decreased DDR1 expression has been associated with the epithelial to mesenchymal transition process in breast cancer, and its overexpression in aggressive basal-like breast cancer cells reduces their invasiveness in 3D cultures and in vivo. In the present work, we propose to study the role of MT1-MMP in the resistance against collagen-induced apoptosis in basal-like breast carcinoma MDA-MB-231 cells. We aimed to investigate whether MT1-MMP depletion is able to restore apoptosis mediated by collagen/DDR1/BIK axis and to verify if such depletion is able to restore full-length DDR1 expression and phosphorylation. ShRNA strategy against MT1-MMP mRNA was able to partially restore full length DDR1 expression and phosphorylation. This was accompanied by a decrease in cell growth and an upregulation of BIK expression. This suggested that MT1-MMP expression in basal-like breast carcinoma cells, in addition to a low basal level of DDR1 expression, protects these cells against collagen-induced apoptosis via DDR1 cleavage. Since DDR1 was moderately expressed in MDA-MB-231 cells, we then investigated whether overexpression of DDR1 could be able to increase its ability to suppress cell growth and to induce apoptosis. Data showed that overexpression of DDR1 induced a decrease in cell growth and an increase in BIK expression, suggesting that moderate expression level of full length DDR1 in basal-like breast carcinoma provides them with a capacity to resist to collagen-induced cell growth suppression and apoptosis. Finally, the combined overexpression of DDR1 and depletion of MT1-MMP in MDA-MB-231 cells synergistically increased collagen-induced cell growth suppression and apoptosis to a level similar to that observed in luminal breast carcinoma. Taken together, our data suggest that during the acquisition of mesenchymal features, the low level of DDR1 expression should be considered as an important biomarker in the prognosis of basal-like breast carcinoma, conferring them a high rate of cell growth and resistance to BIK-mediated apoptosis induced by the stromal collagen.

Pathological and radiological assessments of Paget’s disease

Background: Pure Paget’s disease (PD) of the nipple is a non-invasive breast carcinoma. However, choosing the optimal surgical procedure is difficult due to tumour location and spread into the breast. Thus, in the current study, we retrospectively investigated PD cases in our institution to clarify the details of this disease. We also considered optimal surgical procedures. Methods: Among 4,341 breast cancer cases in the last 11 years at our institution, we retrospectively investigated 17 PD cases with no mass lesion in the breast. Results: Total mastectomy was performed in 15 patients. One patient initially requested central lumpectomy (CL) but the procedure was switched to mastectomy due to a positive surgical margin, demonstrated intraoperatively. On final pathological diagnosis, two patients had PD alone (12%). PD with ductal carcinoma in situ (PD + DCIS) and invasive ductal carcinoma (PD + IDC) were observed in 10 (59%) and 5 (29%) patients, respectively. As to oestrogen receptor (ER) and epidermal growth factor receptor 2 (HER2) statuses, ER-positive/HER2-negative, ER-positive/HER2-positive, ER-negative/HER2-positive and ER-negative/HER2-negative tumours were observed in 6%, 0%, 76% and 18%, respectively, of patients. Disease spread into the breast was pre-operatively suspected in 8 cases, based on micro-calcifications on MMG, and in 8 cases with segmental enhancement on MRI. Sensitivity and specificity of intra-breast disease detection were 62% and 100%, respectively, in 14 patients who had both MMG and MRI findings of PD. Conclusions: Our data confirm that breast conserving surgery is difficult in patients with PD. The surgical procedure might have to be determined in consideration of a balance between curability and cosmesis.

Age-related modifications of type I collagen impair DDR1-induced apoptosis in non-invasive breast carcinoma cells

ABSTRACTType I collagen and DDR1 axis has been described to decrease cell proliferation and to initiate apoptosis in non-invasive breast carcinoma in three-dimensional cell culture matrices. Moreover, MT1-MMP down-regulates these effects. Here, we address the effect of type I collagen aging and MT1-MMP expression on cell proliferation suppression and induced-apoptosis in non-invasive MCF-7 and ZR-75-1 breast carcinoma. We provide evidence for a decrease in cell growth and an increase in apoptosis in the presence of adult collagen when compared to old collagen. This effect involves a differential activation of DDR1, as evidenced by a higher DDR1 phosphorylation level in adult collagen. In adult collagen, inhibition of DDR1 expression and kinase function induced an increase in cell growth to a level similar to that observed in old collagen. The impact of aging on the sensitivity of collagen to MT1-MMP has been reported recently. We used the MT1-MMP expression strategy to verify whether, by degrading adult type I collagen, it could lead to the same phenotype observed in old collagen 3D matrix. MT1-MMP overexpression abrogated the proliferation suppression and induced-apoptosis effects only in the presence of adult collagen. This suggests that differential collagen degradation by MT1-MMP induced a structural disorganization of adult collagen and inhibits DDR1 activation. This could in turn impair DDR1-induced cell growth suppression and apoptosis. Taken together, our data suggest that modifications of collagen structural organization, due to aging, contribute to the loss of the growth suppression and induced apoptosis effect of collagen in luminal breast carcinoma. MT1-MMP-dependent degradation and aging of collagen have no additive effects on these processes.

Abstract B43: Validation of a network-based strategy for the optimization of combinatorial target selection in breast cancer therapy

Abstract Network-based strategies provided by systems biology are attractive tools for cancer therapy. Modulation of cancer networks by anticancer drugs may alter the response of malignant cells and/or drive network reorganization into the inhibition of cancer progression. Previously, using systems biology approach and cancer signaling networks, we identified the top 5 highly expressed and connected proteins (HSP90AB1, CSNK2B, TK1, YWHAB, and VIM) in the invasive MDA-MB-231 breast cancer cell line. Here, we have knocked down the expression of these proteins, individually or together using siRNAs. The transfected cell lines were assessed for in vitro cell growth, colony formation, migration, and invasion relative to control transfected MDA-MB-231, the noninvasive MCF-7 breast carcinoma cell line, and the nontumoral mammary epithelial cell line MCF-10A. The knockdown of the top-5 upregulated connectivity hubs successfully inhibited the in vitro proliferation, colony formation, anchorage independence, migration, and invasion in MDA-MB-231 cells, with minimal effects in the control transfected MDA-MB-231 cells or MCF-7 and MCF-10A cells. The in vitro validation of bioinformatics predictions regarding optimized multitarget selection for therapy suggests that protein expression levels together with protein-protein interaction network analysis may provide an optimized combinatorial target selection for a highly effective antimetastatic precision therapy in triple-negative breast cancer. This approach increases the ability to identify not only druggable hubs as essential targets for cancer survival, but also interactions most susceptible to synergistic drug action. The data provided in this report constitute a preliminary step toward the personalized clinical application of our strategy to optimize the therapeutic use of anticancer drugs. Citation Format: Tatiana Martins Tilli, Nicolas Carels, Jack Adam Tuszynski, Manijeh Pasdar. Validation of a network-based strategy for the optimization of combinatorial target selection in breast cancer therapy [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr B43.

A Case of Non-invasive Intracystic Male Breast Carcinoma

A Case of Non-invasive Intracystic Male Breast Carcinoma

Expression of periostin in breast cancer cells.

Periostin (POSTN) is a protein involved in multiple processes important for cancer development, both at the stage of cancer initiation and progression, as well as metastasis. The aim of this study was to determine the expression of POSTN in the cells of non-invasive ductal breast carcinoma insitu(DCIS) and invasive ductal carcinoma (IDC) and to correlate it with clinicopathological data. Immunohistochemical studies(IHC) were conducted on 21cases of fibrocystic breast change(FC), 44cases of DCIS and 92cases of IDC. POSTN expression at mRNA (real-time PCR) and protein level (western blot analysis) was also confirmed in selected breast cancer cell lines (MCF-7, SK-BR-3, MDA-MB-231 and BO2). Statistically significant higher level of POSTN expression in IDC and DCIS cancer cells compared to FC was noted. Also, the level of POSTN expression in the cytoplasm of IDC cells was shown to increase with the increasing degree of tumour malignancy(G) and significantly higher expression of POSTN was observed in each degree of tumour malignancy(G) relative to FC. Statistically significant higher POSTN expression was observed in tumours with estrogen receptor-negative (ER-) and progesterone receptor-negative (PR-) phenotypes in comparison to estrogen receptor-positive (ER+) and progesterone receptor-positive (PR+) cases, as well as significant negative correlation between POSTN expression in cancer cells and expression of ER and PR (p<0.05). Additionally, statistically significant differences in POSTN expression were shown between particular breast cancer cell lines, both at mRNA and protein level. Observed POSTN expression was the lowest in the case of MCF-7, and the highest in MDA-MB-231 and BO2 of the most aggressive potential clinically corresponding to G3 tumours. POSTN expression in the cytoplasm of IDC cancer cells may play an important role in cancer transformation mechanism.

MiR-130b-3p inhibits cell invasion and migration by targeting the Notch ligand Delta-like 1 in breast carcinoma

Breast carcinoma is the most common malignancy in women, and the incidence rate has increased dramatically in recent years. Metastasis is responsible for most advanced breast cancer mortality, but the underlying mechanisms remain poorly understood despite extensive research. Recently, short non-coding RNA molecules, including miRNAs, which mediate changes in signalling pathways, have emerged as metastatic regulators of the breast carcinoma. Previous reports have suggested that miR-130b-3p has both oncogenic and tumour suppressor functions in a cancer type-dependent manner. However, the roles and underlying molecular mechanisms of miR-130b-3p in the development of metastasis in breast carcinoma remain unclear. Here, we reported for the first time that miR-130b-3p was differentially expressed in early-stage non-invasive MCF-7 human breast carcinoma cells and aggressive late-stage MDA-MB-231 cells. In gain-of-function and loss-of-function studies, we demonstrated that miR-130b-3p could inhibit breast carcinoma cell invasion and migration by directly targeting the Notch ligand Delta-like 1 (DLL1). Our data also indicated that MMP-9, MMP-13, and VEGF were regulated by miR-130b-3p and may be involved in the inhibition of cell invasion and migration in breast carcinoma. Collectively, our findings reveal a new regulatory mechanism of miR-130b-3p and suggest that miR-130b-3p may be a potential target against human breast cancer metastasis.

Validation of a network-based strategy for the optimization of combinatorial target selection in breast cancer therapy: siRNA knockdown of network targets in MDA-MB-231 cells as an in vitro model for inhibition of tumor development.

Network-based strategies provided by systems biology are attractive tools for cancer therapy. Modulation of cancer networks by anticancer drugs may alter the response of malignant cells and/or drive network re-organization into the inhibition of cancer progression. Previously, using systems biology approach and cancer signaling networks, we identified top-5 highly expressed and connected proteins (HSP90AB1, CSNK2B, TK1, YWHAB and VIM) in the invasive MDA-MB-231 breast cancer cell line. Here, we have knocked down the expression of these proteins, individually or together using siRNAs. The transfected cell lines were assessed for in vitro cell growth, colony formation, migration and invasion relative to control transfected MDA-MB-231, the non-invasive MCF-7 breast carcinoma cell line and the non-tumoral mammary epithelial cell line MCF-10A. The knockdown of the top-5 upregulated connectivity hubs successfully inhibited the in vitro proliferation, colony formation, anchorage independence, migration and invasion in MDA-MB-231 cells; with minimal effects in the control transfected MDA-MB-231 cells or MCF-7 and MCF-10A cells. The in vitro validation of bioinformatics predictions regarding optimized multi-target selection for therapy suggests that protein expression levels together with protein-protein interaction network analysis may provide an optimized combinatorial target selection for a highly effective anti-metastatic precision therapy in triple-negative breast cancer. This approach increases the ability to identify not only druggable hubs as essential targets for cancer survival, but also interactions most susceptible to synergistic drug action. The data provided in this report constitute a preliminary step toward the personalized clinical application of our strategy to optimize the therapeutic use of anti-cancer drugs.

MiR-155 expression level changes might be associated with initial phases of breast cancer pathogenesis and lymph-node metastasis.

Breast carcinoma is heterogeneous disease. Understanding the process of invasion and metastasis and the selection of the therapy for patients with breast carcinomas still remains difficult. MicroRNAs are powerful gene expression regulators. Because of inconsistent findings, we have analyzed potential difference in miR-155 levels in three breast cancer groups. Our goals were to examine miR-155 expression levels in normal tissue, non-invasive and invasive breast carcinomas, and their association with standard clinical and pathological parameters and oncomiR-21, and to investigate the ability of miR-155 to separate invasive breast carcinomas with non-invasive component from pure invasive. In the group of 40 breast tissue samples, relative expression levels of miR-155 were examined with stem-loop quantitative real-time PCR using TaqMan technology. The significant difference among four examined groups of the breast tissue was detected (p = 0.001). In the group of pure invasive tumors, patients with positive nodal status had significantly higher miR-155 levels (p = 0.046). Our results suggest that miR-155 might be involved in breast cancer pathogenesis and in tumor spreading to the lymph nodes, and that it might be used as biomarker for additional stratification of patients with invasive breast carcinomas with non-invasive component.

Evaluation of Apparent Diffusion Coefficient to Predict Grade, Microinvasion, and Invasion in Ductal Carcinoma In Situ of the Breast

To evaluate the role of apparent diffusion coefficient (ADC) in distinguishing ductal carcinoma in situ (DCIS) grades and identifying microinvasive and/or invasive disease in the preoperative evaluation of patients with core biopsy-proven DCIS. Research Ethics Board-approved study with informed consent from 81 women (age, 36-84 years) scheduled for core-biopsy with results of 82 noninvasive breast carcinomas. All patients were assessed preoperatively by diffusion sequence in addition to contrast magnetic resonance imaging (MRI). Lesion morphology and ADC values were recorded. The Kruskal-Wallis or one-way analysis of variance test and Pearson correlation coefficient were used to study the association between ADC and MRI lesion characteristics. Logistic regression analysis was used to evaluate the ability of ADC to predict the presence of invasion. Surgical pathology demonstrated associated invasive cancer in 26.8%, microinvasion in 14.6%, and pure DCIS in 58.5%. The minimum regions of interest (ROI)-based ADC was significantly different among the following three groups (P < .001, Kruskal-Wallis test): 0.98 × 10(-3) mm(2)/s ± 0.25 for pure DCIS, 0.82 × 10(-3) mm(2)/s ± 0.20 for DCIS with microinvasion, and 0.71 × 10(-3) mm(2)/s ± 0.27 for DCIS with invasive disease. Based on logistic regression analysis, the minimum ROI-based ADC of 0.56 × 10(-3) mm(2)/s was a significant predictor for invasive disease (odds ratio = 0.02, 95% confidence interval [0.002, 0.207], P = .001). Regardless of the field strength (1.5 vs. 3.0 T) ADC values of high-grade and non-high-grade DCIS were not significantly different. Pure DCIS had the highest "ROI-based" ADC measured using 1.5 T or 3.0 T. The ADC was able to identify microinvasion or invasive cancer in biopsy-proven DCIS lesions but not to distinguish the DCIS grades.

男性非浸潤性嚢胞内乳癌の1例

男性乳癌は，比較的稀な疾患で全乳癌症例に占める割合は0.5～1.0％とされ，中でも非浸潤性嚢胞内癌はごく稀である．症例は76歳の男性，左乳房腫瘤を主訴に来院．視触診で左乳輪から頭側にわたる領域に2cmの境界明瞭な腫瘤と軽度陥没乳頭を認め，腋窩リンパ節の腫脹は認めなかった．マンモグラフィーで境界明瞭腫瘤陰影を認め（カテゴリー3），乳房超音波検査では嚢胞内に立ち上がりのなだらかな腫瘤を認めカテゴリー4（嚢胞内癌の疑い）と診断した．前医での細胞診ではclass III，針生検で非浸潤性乳管癌もしくは浸潤癌の疑いと診断された．乳房部分切除を施行し，最終病理診断は非浸潤性嚢胞内癌であった．残存乳腺に放射線照射（50Gy）を行い，経過は良好である．

The difference in miR-21 expression levels between invasive and non-invasive breast cancers emphasizes its role in breast cancer invasion

MicroRNA-21 (miR-21) overexpression is characteristic for various types of tumors, but it is still unknown whether its expression levels differ between invasive and non-invasive breast carcinomas. The main goal of the study was to determine the difference in miR-21 expression among normal tissue, non-invasive, invasive with non-invasive component, and pure invasive breast cancer samples, to explain its potential role and significance in breast cancer invasiveness. The second goal was to propose miR-21 as molecular marker of breast cancer invasiveness and potential target for future anti-miR therapies for the prevention of invasion and metastasis. In order to reveal the role of miR-21 in breast cancer invasiveness, we measured miR-21 expression levels in 44 breast cancer and four normal samples by stem-loop real-time RT-PCR using TaqMan technology. Relative expression levels of miR-21 were significantly higher in invasive than in other groups (P=0.002) and significantly higher in invasive compared with invasive with non-invasive component group in histological (P=0.043) and nuclear grade 2 (P=0.036), estrogen-receptor-positive (ER+) (P=0.006), progesterone-receptor-positive (PR+) (P=0.008), ER+PR+ (P=0.007), and proliferation index (Ki-67)≤20% (P=0.036) tumors. Our findings suggest that miR-21 could be independent molecular marker of breast cancer invasiveness and potential target for future anti-miR therapies for the prevention of invasion and metastasis.