Abstract:Infiltrating lobular breast cancer (ILC) is the most common special breast cancer subtype. With mutational or epigenetic inactivation of the cell adhesion molecule E-cadherin (CDH1) being confined almost exclusively to ILC, this tumor entity stands out from all other types of breast cancers. The molecular basis of ILC is linked to loss of E-cadherin, as evidenced by human CDH1 germline mutations and conditional knockout mouse models. A better understanding of ILC beyond the level of descriptive studies depends… Show more
“…Only BCK4 and T47D had detectable PR expression at these exposure conditions. As expected, E-cadherin was absent from MDA-MB-134, SUM44, which harbor CDH1 truncating mutations (30), from BCK4 cells, as well as from MDA-MB-231 cells, in which the CDH1 promoter is hypermethylated (31). Figure 1.Characteristics of the human ILC and IDC cell lines used in the study. A.…”
Section: Resultssupporting
confidence: 70%
“…1A , Western blotting confirmed ER expression in all cell lines, except for the ER-negative MDA-MB-231 cells (29). Of note, although there are conflicting reports on the ER status of MDA-MB-330 cells (29,30), they displayed abundant ER levels in our hands. Only BCK4 and T47D had detectable PR expression at these exposure conditions.…”
Section: Resultscontrasting
confidence: 60%
“…Consistent with the down-regulation and/or mislocalization of other junction components in the absence of E-cadherin (32,33), β-catenin expression was absent from MDA-MB-134, SUM44 and BCK4 cells – a result different from that in MDA-MB-231 cells, which retained β-catenin expression without E-cadherin. Interestingly, MDA-MB-330 cells, which harbor a bi-allelic, truncating CTNNA1 mutation (30), still expressed E-cadherin and β-catenin in the absence of α-catenin. In contrast, p120-catenin (p120) was detected in all cell lines with the weakest expression in BCK4 cells, which also exhibited lower α-catenin levels.…”
250 words) 83 84 Invasive lobular carcinoma (ILC) is the second most common subtype of breast cancer following 85 invasive ductal carcinoma (IDC) and characterized by the loss of E-cadherin-mediated adherens 86 junctions. Despite displaying unique histological and clinical features, ILC still remains a 87 chronically understudied disease with limited knowledge on the available laboratory research 88 models. To this end, herein we report a comprehensive 2D and 3D phenotypic characterization of 89 four Estrogen Receptor-positive human ILC cell lines -MDA-MB-134, SUM44, MDA-MB-330 90 and BCK4. Compared to the IDC cell lines MCF7, T47D and MDA-MB-231, ultra-low 91 attachment culture conditions revealed a remarkable anchorage-independence ability that was 92 unique to the ILC cells, a feature not evident in soft agar gels. 3D Collagen I and Matrigel 93 culture indicated a generally loose morphology for the ILC cell lines, which exhibited differing 94 preferences for adhesion to ECM proteins in 2D. Furthermore, ILC cells had limited migration 95 and invasion ability in wound-scratch and transwell assays with the exception of haptotaxis to 96 Collagen I. Transcriptional comparison of the cell lines confirmed the decreased cell 97 proliferation and E-cadherin-mediated intercellular junctions in ILC, while uncovering the 98 induction of novel pathways related to cyclic nucleotide phosphodiesterase activity, ion 99 channels, drug metabolism and alternative cell adhesion molecules such as N-cadherin, some of 100 which were also differentially regulated in ILC versus IDC tumors. Altogether, these studies will 101 serve as an invaluable resource for the breast cancer research community and facilitate further 102 functional discoveries towards understanding ILC, identifying novel drug targets and ultimately 103 improving the outcome of patients with ILC.
“…Only BCK4 and T47D had detectable PR expression at these exposure conditions. As expected, E-cadherin was absent from MDA-MB-134, SUM44, which harbor CDH1 truncating mutations (30), from BCK4 cells, as well as from MDA-MB-231 cells, in which the CDH1 promoter is hypermethylated (31). Figure 1.Characteristics of the human ILC and IDC cell lines used in the study. A.…”
Section: Resultssupporting
confidence: 70%
“…1A , Western blotting confirmed ER expression in all cell lines, except for the ER-negative MDA-MB-231 cells (29). Of note, although there are conflicting reports on the ER status of MDA-MB-330 cells (29,30), they displayed abundant ER levels in our hands. Only BCK4 and T47D had detectable PR expression at these exposure conditions.…”
Section: Resultscontrasting
confidence: 60%
“…Consistent with the down-regulation and/or mislocalization of other junction components in the absence of E-cadherin (32,33), β-catenin expression was absent from MDA-MB-134, SUM44 and BCK4 cells – a result different from that in MDA-MB-231 cells, which retained β-catenin expression without E-cadherin. Interestingly, MDA-MB-330 cells, which harbor a bi-allelic, truncating CTNNA1 mutation (30), still expressed E-cadherin and β-catenin in the absence of α-catenin. In contrast, p120-catenin (p120) was detected in all cell lines with the weakest expression in BCK4 cells, which also exhibited lower α-catenin levels.…”
250 words) 83 84 Invasive lobular carcinoma (ILC) is the second most common subtype of breast cancer following 85 invasive ductal carcinoma (IDC) and characterized by the loss of E-cadherin-mediated adherens 86 junctions. Despite displaying unique histological and clinical features, ILC still remains a 87 chronically understudied disease with limited knowledge on the available laboratory research 88 models. To this end, herein we report a comprehensive 2D and 3D phenotypic characterization of 89 four Estrogen Receptor-positive human ILC cell lines -MDA-MB-134, SUM44, MDA-MB-330 90 and BCK4. Compared to the IDC cell lines MCF7, T47D and MDA-MB-231, ultra-low 91 attachment culture conditions revealed a remarkable anchorage-independence ability that was 92 unique to the ILC cells, a feature not evident in soft agar gels. 3D Collagen I and Matrigel 93 culture indicated a generally loose morphology for the ILC cell lines, which exhibited differing 94 preferences for adhesion to ECM proteins in 2D. Furthermore, ILC cells had limited migration 95 and invasion ability in wound-scratch and transwell assays with the exception of haptotaxis to 96 Collagen I. Transcriptional comparison of the cell lines confirmed the decreased cell 97 proliferation and E-cadherin-mediated intercellular junctions in ILC, while uncovering the 98 induction of novel pathways related to cyclic nucleotide phosphodiesterase activity, ion 99 channels, drug metabolism and alternative cell adhesion molecules such as N-cadherin, some of 100 which were also differentially regulated in ILC versus IDC tumors. Altogether, these studies will 101 serve as an invaluable resource for the breast cancer research community and facilitate further 102 functional discoveries towards understanding ILC, identifying novel drug targets and ultimately 103 improving the outcome of patients with ILC.
“…CRISPR-Cas9-mediated CDH1 KO is induced by a single base pair deletion, which generated a premature stop codon, mimicking missense mutaitons found in ILC tumors and in genetically characterized ILC cell lines 30 . This point mutation led to depletion of both E-cadherin RNA and protein ( Fig.…”
Section: Cdh1 Point Mutation Results In Loss Of Spliced E-cadherin Rnmentioning
Invasive lobular breast carcinoma (ILC), one of the major breast cancer histological subtypes, exhibits unique clinical and molecular features compared to the other well-studied ductal cancer subtype (IDC). The pathognomonic feature of ILC is loss of E-cadherin, mainly caused by inactivating mutations within the CDH1 gene, but the extent of contribution of this genetic alteration to ILC-specific molecular characteristics remains largely understudied. To profile these features transcriptionally, we conducted single cell RNA sequencing on a panel of IDC and ILC cell lines, as well as an IDC cell line (T47D) with CRISPR-Cas9-mediated knock out (KO) of CDH1. Inspection of intra-cell line heterogeneity illustrated genetically and transcriptionally distinct subpopulations in multiple cell lines and highlighted rare populations of MCF7 cells highly expressing an apoptosis-related signature, positively correlated with a pre-adaptation signature to estrogen deprivation. Investigation of CDH1 KO-induced alterations showed transcriptomic membranous systems remodeling, elevated resemblance to ILCs in regulon activation, and suggests IRF1 as a potential mediator of reduced proliferation and increased cytokine-mediated immune-reactivity in ILCs.
“…In breast cancer, mode and timing of E-cadherin inactivation appears to determine tumor type and etiology [2]. In diffuse gastric cancer and invasive lobular breast cancer (ILC), E-cadherin loss is an early and causative lesion [3–7], while most other tumors show loss of E-cadherin during later stages of disease progression (reviewed in: [8]).…”
Loss of E-cadherin expression is causal to the development of invasive lobular breast carcinoma (ILC). E-cadherin loss leads to dismantling of the adherens junction and subsequent translocation of p120-catenin (p120) to the cytosol and nucleus. Although p120 is critical for the metastatic potential of ILC through the regulation of Rock-dependent anoikis resistance, it remains unknown whether p120 also contributes to ILC development. Using genetically engineered mouse models with mammary gland-specific inactivation of E-cadherin, p120 and p53, we demonstrate that ILC formation induced by E-cadherin and p53 loss is severely impaired upon concomitant inactivation of p120. Tumors that developed in the triple-knockout mice were mostly basal sarcomatoid carcinomas that displayed overt nuclear atypia and multinucleation. In line with the strong reduction in ILC incidence in triple-knockout mice compared to E-cadherin and p53 double-knockout mice, no functional redundancy of p120 family members was observed in mouse ILC development, as expression and localization of ARVCF, p0071 or δ-catenin was unaltered in ILCs from triple-knockout mice. In conclusion, we show that loss of p120 in the context of the p53-deficient mouse models is dominant over E-cadherin inactivation and its inactivation promotes the development of basal, epithelial-to-mesenchymal-transition (EMT)-type invasive mammary tumors.Electronic supplementary materialThe online version of this article (doi:10.1007/s10911-016-9358-3) contains supplementary material, which is available to authorized users.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
