scholarly journals Novel Image Analysis Approach Quantifies Morphological Characteristics of 3D Breast Culture Acini with Varying Metastatic Potentials

2012 ◽  
Vol 2012 ◽  
pp. 1-16 ◽  
Author(s):  
Lindsey McKeen Polizzotti ◽  
Basak Oztan ◽  
Chris S. Bjornsson ◽  
Katherine R. Shubert ◽  
Bülent Yener ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Image Analysis ◽  
Three Dimensional ◽  
Morphological Characteristics ◽  
Tissue Samples ◽  
Metastatic Cascade ◽  
Automated Grading

Prognosis of breast cancer is primarily predicted by the histological grading of the tumor, where pathologists manually evaluate microscopic characteristics of the tissue. This labor intensive process suffers from intra- and inter-observer variations; thus, computer-aided systems that accomplish this assessment automatically are in high demand. We address this by developing an image analysis framework for the automated grading of breast cancer inin vitrothree-dimensional breast epithelial acini through the characterization of acinar structure morphology. A set of statistically significant features for the characterization of acini morphology are exploited for the automated grading of six (MCF10 series) cell line cultures mimicking three grades of breast cancer along the metastatic cascade. In addition to capturing both expected and visually differentiable changes, we quantify subtle differences that pose a challenge to assess through microscopic inspection. Our method achieves 89.0% accuracy in grading the acinar structures as nonmalignant, noninvasive carcinoma, and invasive carcinoma grades. We further demonstrate that the proposed methodology can be successfully applied for the grading ofin vivotissue samples albeit with additional constraints. These results indicate that the proposed features can be used to describe the relationship between the acini morphology and cellular function along the metastatic cascade.

scholarly journals Splicing factor SRSF1 promotes breast cancer progression via oncogenic splice switching of PTPMT1

2021 ◽  
Vol 40 (1) ◽  
Author(s):  
Jun-Xian Du ◽  
Yi-Hong Luo ◽  
Si-Jia Zhang ◽  
Biao Wang ◽  
Cong Chen ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Progression ◽  
High Risk Group ◽  
Clinical Samples ◽  
Binding Motif ◽  
Ki 67 ◽  
Tissue Samples

Abstract Background Intensive evidence has highlighted the effect of aberrant alternative splicing (AS) events on cancer progression when triggered by dysregulation of the SR protein family. Nonetheless, the underlying mechanism in breast cancer (BRCA) remains elusive. Here we sought to explore the molecular function of SRSF1 and identify the key AS events regulated by SRSF1 in BRCA. Methods We conducted a comprehensive analysis of the expression and clinical correlation of SRSF1 in BRCA based on the TCGA dataset, Metabric database and clinical tissue samples. Functional analysis of SRSF1 in BRCA was conducted in vitro and in vivo. SRSF1-mediated AS events and their binding motifs were identified by RNA-seq, RNA immunoprecipitation-PCR (RIP-PCR) and in vivo crosslinking followed by immunoprecipitation (CLIP), which was further validated by the minigene reporter assay. PTPMT1 exon 3 (E3) AS was identified to partially mediate the oncogenic role of SRSF1 by the P-AKT/C-MYC axis. Finally, the expression and clinical significance of these AS events were validated in clinical samples and using the TCGA database. Results SRSF1 expression was consistently upregulated in BRCA samples, positively associated with tumor grade and the Ki-67 index, and correlated with poor prognosis in a hormone receptor-positive (HR+) cohort, which facilitated proliferation, cell migration and inhibited apoptosis in vitro and in vivo. We identified SRSF1-mediated AS events and discovered the SRSF1 binding motif in the regulation of splice switching of PTPMT1. Furthermore, PTPMT1 splice switching was regulated by SRSF1 by binding directly to its motif in E3 which partially mediated the oncogenic role of SRSF1 by the AKT/C-MYC axis. Additionally, PTPMT1 splice switching was validated in tissue samples of BRCA patients and using the TCGA database. The high-risk group, identified by AS of PTPMT1 and expression of SRSF1, possessed poorer prognosis in the stage I/II TCGA BRCA cohort. Conclusions SRSF1 exerts oncogenic roles in BRCA partially by regulating the AS of PTPMT1, which could be a therapeutic target candidate in BRCA and a prognostic factor in HR+ BRCA patient.

Prevascularized, Co-Culture Model for Breast Cancer Drug Development

2012 ◽  
Author(s):  
Lauren Marshall ◽  
Isabel Löwstedt ◽  
Paul Gatenholm ◽  
Joel Berry
Keyword(s):  
Breast Cancer ◽  
Drug Development ◽  
Three Dimensional ◽  
Human Tumor ◽  
3D Models ◽  
Cancer Drug ◽  
Cancer Therapies ◽  
Anti Cancer

The objective of this study was to create 3D engineered tissue models to accelerate identification of safe and efficacious breast cancer drug therapies. It is expected that this platform will dramatically reduce the time and costs associated with development and regulatory approval of anti-cancer therapies, currently a multi-billion dollar endeavor [1]. Existing two-dimensional (2D) in vitro and in vivo animal studies required for identification of effective cancer therapies account for much of the high costs of anti-cancer medications and health insurance premiums borne by patients, many of whom cannot afford it. An emerging paradigm in pharmaceutical drug development is the use of three-dimensional (3D) cell/biomaterial models that will accurately screen novel therapeutic compounds, repurpose existing compounds and terminate ineffective ones. In particular, identification of effective chemotherapies for breast cancer are anticipated to occur more quickly in 3D in vitro models than 2D in vitro environments and in vivo animal models, neither of which accurately mimic natural human tumor environments [2]. Moreover, these 3D models can be multi-cellular and designed with extracellular matrix (ECM) function and mechanical properties similar to that of natural in vivo cancer environments [3].

Characterization of Folic Acid Surface-Coated Selenium Nanoparticles and Corresponding In Vitro and In Vivo Effects Against Breast Cancer

2018 ◽  
Vol 49 (1) ◽  
pp. 10-17 ◽  
Author(s):  
Ahmad Reza Shahverdi ◽  
Faranak Shahverdi ◽  
Elnaz Faghfuri ◽  
Mohammad Reza khoshayand ◽  
Faranak Mavandadnejad ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Folic Acid ◽  
Selenium Nanoparticles ◽  
In Vivo Effects

Primary breast cancer tryptase expression as novel molecular drug target.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. e21113-e21113
Author(s):  
Girolamo Ranieri ◽  
Michele Ammendola ◽  
Mario Brandi ◽  
Andrea Misino ◽  
Veronica Goffredo ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Image Analysis ◽  
Mast Cells ◽  
Vascular Endothelial Cells ◽  
Receptor Activation ◽  
Female Breast Cancer ◽  
Serial Sections ◽  
Kit Receptor

e21113 Background: Tryptase, a serine protease stored and released from mast cells granules has been identified as a new non-classical angiogenetic factor. Mast cells can release tryptase following c-Kit receptor activation. We have evaluated the correlations among the number of MCs positive to tryptase (MCDPT), the number of c-Kit receptor expressing cells (C-KREC) and microvascular density (MVD) in a series of 88 primary T1-3, N0-2 M0 female breast cancer by means of immunohistochemistry and image analysis methods. Methods: Six-micrometers thick serial sections of formalin-fixed and paraffin-embedded bioptic tumor samples were microwaved at 500 W for 10 min. and treated with a 3% hydrogen peroxide solution. Sections were incubated with primary antibodies: anti-tryptase (AA1; Dako, Glostrup, Denmark), anti-c-Kit receptor (A4502;Dako, Glostrup, Denmark) and anti-CD34 (QB-END 10; Bio-Optica Milan, Italy). In serial sections MVD, MCDPT and C-KREC were counted by means of image analysis at x400. Results: Data demonstrated a significantly (r= ranging from 0.71 to 0.91; p: ranging from 0.001 to 0.003 by Pearson’s analysis respectively) correlation between MVD, MCDPT and C-KREC to each other. Conclusions: Published in vitro data suggest that tryptase induce angiogenesis in vascular endothelial cells and breast cancer cells lines. According to these data we shown that MVD, MCDPT and C-KREC paralleled to each other suggesting a role in in vivo breast cancer angiogenesis. In this context several tryptase inhibitors such as gabexate mesilate or nafamostat mesilate might be evaluated in clinical trials as a new antiangiogenetic approach.

Emerging Vascular Applications of Magnetic Resonance Imaging: A Picture is Worth More than a Thousand Words

Vascular ◽  
2006 ◽  
Vol 14 (6) ◽  
pp. 366-371 ◽  
Author(s):  
Tamara N. Fitzgerald ◽  
Akihito Muto ◽  
Fabio Akimaro Kudo ◽  
Jose Mario Pimiento ◽  
Robert Todd Constable ◽  
...  
Keyword(s):  
Magnetic Resonance ◽  
Three Dimensional ◽  
Quantitative Information ◽  
Data Set ◽  
Vascular Intervention ◽  
In Vivo Experiments ◽  
Blood Flow Dynamics

Vascular applications of magnetic resonance (MR) imaging are reviewed, with emphasis on algorithms that use nonpictorial information contained in the MR data set. Current clinical vascular practice generally limits use of MR angiography and three-dimensional vessel images to qualitative pictorial rendering without routinely using the available quantitative information contained within the MR data. This review is dedicated to recent advances that include characterization of vessel histology, assessment of carotid plaque vulnerability, characterization of blood flow dynamics, quantitative analysis of disease severity, and prediction of vascular intervention outcome. Examples from histologic preparation, in vitro and in vivo experiments, are discussed, with an emphasis on potential clinical applications and advances in acquisition technology.

scholarly journals Parvin-β Inhibits Breast Cancer Tumorigenicity and Promotes CDK9-Mediated Peroxisome Proliferator-Activated Receptor Gamma 1 Phosphorylation

2007 ◽  
Vol 28 (2) ◽  
pp. 687-704 ◽  
Author(s):  
Cameron N. Johnstone ◽  
Perry S. Mongroo ◽  
A. Sophie Rich ◽  
Michael Schupp ◽  
Mark J. Bowser ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Cancer Progression ◽  
Breast Cancer Cells ◽  
Three Dimensional ◽  
Nuclear Hormone Receptor ◽  
Peroxisome Proliferator ◽  
Peroxisome Proliferator Activated Receptor

ABSTRACT Parvin-β is a focal adhesion protein downregulated in human breast cancer cells. Loss of Parvin-β contributes to increased integrin-linked kinase activity, cell-matrix adhesion, and invasion through the extracellular matrix in vitro. The effect of ectopic Parvin-β expression on the transcriptional profile of MDA-MB-231 breast cancer cells, which normally do not express Parvin-β, was evaluated. Particular emphasis was placed upon propagating MDA-MB-231 breast cancer cells in three-dimensional culture matrices. Interestingly, Parvin-β reexpression in MDA-MB-231 cells increased the mRNA expression, serine 82 phosphorylation (mediated by CDK9), and activity of the nuclear hormone receptor peroxisome proliferator-activated receptor gamma (PPARγ), and there was a concomitant increase in lipogenic gene expression as a downstream effector of PPARγ. Importantly, Parvin-β suppressed breast cancer growth in vivo, with associated decreased proliferation. These data suggest that Parvin-β might influence breast cancer progression.

scholarly journals Characterization of Fibroblasts with Son of Sevenless-1 Mutation

2006 ◽  
Vol 85 (11) ◽  
pp. 1050-1055 ◽  
Author(s):  
E.J. Lee ◽  
S.I. Jang ◽  
D. Pallos ◽  
J. Kather ◽  
T.C. Hart
Keyword(s):  
Cell Proliferation ◽  
Three Dimensional ◽  
Collagen Matrix ◽  
Histological Assessment ◽  
M Phase ◽  
Son Of Sevenless ◽  
Gingival Fibromatosis

Although non-syndromic hereditary gingival fibromatosis (HGF) is genetically heterogeneous, etiologic mutations have been identified only in the Son of Sevenless-1 gene ( SOS1). To test evidence of increased cell proliferation, we studied histological, morphological, and proliferation characteristics in monolayer and three-dimensional cultures of fibroblasts with the SOS1 g.126,142–126,143insC mutation. Histological assessment of HGF gingiva indicated increased numbers of fibroblasts (30%) and increased collagen (10%). Cell proliferation studies demonstrated increased growth rates and 5-bromo-2-deoxyuridine incorporation for HGF fibroblasts. Flow cytometry showed greater proportions of HGF fibroblasts in the G2/M phase. Attachment of HGF fibroblasts to different extracellular matrix surfaces demonstrated increased formation of protrusions with lamellipodia. HGF fibroblasts in three-dimensional culture showed greater cell proliferation, higher cell density, and alteration of surrounding collagen matrix. These findings revealed that increased fibroblast numbers and collagen matrix changes are associated with mutation of the SOS1 gene in vitro and in vivo.

scholarly journals Modelling and Validating Three-Dimensional Human Breast and Cancerous Human Breast Tissues In Vitro

2020 ◽  
pp. 1-9
Author(s):  
Anna Karolina Zuk ◽  
Anna Karolina Zuk ◽  
Beata Burczynska ◽  
Dong Li ◽  
Lucy Ghali ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Smooth Muscle Actin ◽  
Three Dimensional ◽  
Myoepithelial Cells ◽  
In Vitro Models ◽  
Normal Breast ◽  
Human Breast ◽  
Mucin 1

In this study three dimensional (3-D) in vitro models of normal breast and breast cancer tissues were developed to mimic closely the in vivo tissue microenvironment and therefore providing reliable models for in vitro studies as well as testing of novel cancer therapies. Normal and cancerous human breast cell lines were used to construct 3-D artificial tissues, where de-epidermalised dermis (DED) was used as a scaffold for both models. Morphological analyses were conducted using haematoxylin and eosin staining. Biomarkers including keratin 5 and 19 as well as α smooth muscle actin and mucin 1 were used to confirm and validate the reliability of the proposed models using immunohistochemical techniques. Findings suggest that the 3-D in vitro models described in this work can serve as functional models of both human normal and cancerous breast tissues. Multiple structures similar to ducts and lobules of human breast in vivo were observed in 3-D in vitro models by the use of H&E, some breast cancer colonies seen in the cancerous 3-D model were similar to the ducto-lobular structures observed in normal 3-D model of the breast but the former cells were more loosely connected, irregular and largely disorganized. The established 3-D in vitro model of normal breast showed the development of ducto-lobular structures composed of an inner cell layer which was stained positive with α mucin 1 antibody, a biomarker that is characteristic for luminal cells; and also an outer basal layer of cells that was stained positive for α smooth muscle actin, a biomarker of myoepithelial cells.. Keratin staining in 3-D in vitro models also resembled the pattern observed in vivo where keratin 5 was detected in both luminal and myoepithelial cells of normal breast model (NTERT cells), whereas keratin 19 was present in breast cancer model (C2321 cells). These 3-D models successfully recapitulate both normal and pathological tissue architecture of breast tissue and has the potential for various applications in the evaluation of breast cancer progression and treatment.

scholarly journals Isolation and characterization of breast cancer stem cells

2019 ◽  
Author(s):  
Ιωάννης Βερίγος
Keyword(s):  
Breast Cancer ◽  
Stem Cells ◽  
Cancer Stem Cells ◽  
Breast Cancer Stem Cells ◽  
Isolation And Characterization

Τα καρκινικά βλαστικά κύτταρα (ΚΒΚ) αποτελούν έναν υποπληθυσμό των καρκινικών κυττάρων με ιδιότητες αυτο-ανανέωσης και διαφοροποίησης, που θεωρείται ότι ευθύνονται για την ανάπτυξη του όγκου και την υποτροπή μετά από θεραπεία. Η ικανότητα των ΚΒΚ να επιβιώνουν μετά από εφαρμογή συμβατικών μορφών αντικαρκινικής θεραπείας συνέβαλε στην προσπάθεια ανάπτυξης νέων θεραπευτικών στρατηγικών που στοχεύουν τα συγκεκριμένα κύτταρα. Πρόσφατες μελέτες, υποδεικνύουν ότι οι μοναδικές ιδιότητες των ΚΒΚ φαίνεται να υπόκεινται σε επιγενετική ρύθμιση. Η LSD1 / KDM1A, μία απομεθυλάση ιστονών, ρυθμίζει τις βλαστικές ιδιότητες των κυττάρων, ενώ επίσης υπερεκφράζεται σε διαφορετικούς τύπους καρκίνου συμπεριλαμβανομένου και του καρκίνου του μαστού. Στην παρούσα εργασία, στόχος μας ήταν να διερευνήσουμε τον ρόλο της LSD1 στην χημειο-ανθεκτικότητα των ΚΒΚ του μαστού. Πειράματα ανοσοϊστοχημείας σε δείγματα ασθενών με καρκίνο του μαστού επιβεβαίωσαν τη σύνδεση της έκφρασης της LSD1 με λιγότερο διαφοροποιημένους όγκους του μαστού. Επιπλέον, ο γονιδιακός χειρισμός της έκφρασης της LSD1 με διαφορετικές πειραματικές προσεγγίσεις, συσχέτισε την απομεθυλάση με την χημειο-ανθεκτικότητα των καρκινικών κυττάρων του μαστού. Επιπροσθέτως, έχοντας εγκαθιδρύσει και χαρακτηρίσει εκτενώς ένα in vitro σύστημα καλλιέργειας σφαιρικών δομών εμπλουτισμένο σε καρκινικά βλαστικά κύτταρα του μαστού, πειράματα γονιδιακής αποσιώπησης και υπερέκφρασης, έδειξαν ότι το ένζυμο ρυθμίζει τις βλαστικές ιδιότητες του υπο-πληθυσμού των καρκινικών βλαστικών κυττάρων του μαστού CD44+/CD24-/low.. Επιπλέον, η φαρμακολογική αναστολή της LSD1 οδήγησε σε μείωση των ΚΒΚ και εξασθένιση του δυναμικού βλαστικότητας in vitro, ενώ ήταν σε θέση να οδηγήσει σε συρρίκνωση των όγκων και να μειώσει τον υποπληθυσμό ΚΒΚ in vivo. Εν κατακλείδι, η ανθεκτικότητα στη χημειοθεραπεία των καρκινικών βλαστικών κυττάρων του μαστού επιβεβαιώθηκε στο σύστημά μας μέσω των αντίστοιχων πειραματικών προσεγγίσεων. Ωστόσο, η χορήγηση ενός αναστολέα της LSD1 κατέστησε τα καρκινικά βλαστικά κύτταρα του μαστού πιο ευαίσθητα στις ευρέως χρησιμοποιούμενες θεραπευτικές μεθόδους, όπως δείχνουν οι ποσοτικοί προσδιορισμοί του ρυθμού πολλαπλασιασμού των κυττάρων και σχηματισμού σφαιρικών δομών. Πειράματα με ξενομοσχεύματα σε επίμυες βρίσκονται σε εξέλιξη για να επιβεβαιωθεί ότι η στόχευση της LSD1 σε συνδυασμό με χημειοθεραπευτικούς παράγοντες αναστέλλει αποτελεσματικότερα την ανάπτυξη όγκων. Τα παραπάνω δεδομένα υποδηλώνουν έντονα ότι η LSD1 αποτελεί έναν πιθανό θεραπευτικό στόχο στα καρκινικά βλαστικά κύτταρα του μαστού, καθώς βρέθηκε ότι ρυθμίζει τις βλαστικές ιδιότητες των κυττάρων αυτών και έτσι προσδίδει χημειο-ανθεκτικές ικανότητες.

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