Detection of bone marrow‐disseminated breast cancer cells using an RT‐PCR assay of MUC5B mRNA

Berois, Nora; Varangot, Mario; Sóñora, Cecilia; Zarantonelli, Leticia; Pressa, Carlos; Laviña, Raúl; Rodrı́guez, José Luis; Delgado, Fernando; Porchet, Nicole; Aubert, Jean‐Pierre; Osinaga, Eduardo

doi:10.1002/ijc.10853

Cited by 31 publications

(24 citation statements)

References 41 publications

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“…Figure 4 shows the expression patterns of a hypothetical protein in this group, FLJ20406. FLJ20406 is predicted to contain a signal sequence and has homology to the mouse Lck-interacting transmembrane adaptor protein (41). Two genes COL10A1 (collagen X, alpha 1) and CLECSF5 (C-type lectin superfamily member 5) were overexpressed in almost all of the cancers tested, although varying levels of expression were observed (Fig.…”

Section: -E Immunohistostaining Confirmed the Pcr Results And Showmentioning

confidence: 99%

Selective Identification of Secreted and Transmembrane Breast Cancer Markers using Escherichia coli Ampicillin Secretion Trap

Ferguson¹,

Muenster²,

Zang³

et al. 2005

Cancer Research

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Secreted and cell surface proteins play important roles in cancer and are potential drug targets and tumor markers. Here, we describe a large-scale analysis of the genes encoding secreted and cell surface proteins in breast cancer. To identify these genes, we developed a novel signal sequence trap method called Escherichia coli ampicillin secretion trap (CAST). For CAST, we constructed a plasmid in which the signal sequence of b-lactamase was deleted such that it does not confer ampicillin resistance. Eukaryotic cDNA libraries cloned into pCAST produced tens of thousands of ampicillinresistant clones, 80% of which contained cDNA fragments encoding secreted and membrane spanning proteins. We identified 2,708 unique sequences from cDNA libraries made from surgical breast cancer specimens. We analyzed the expression of 1,287 of the 2,708 genes and found that 166 were overexpressed in breast cancers relative to normal breast tissues. Eighty-five percent of these genes had not been previously identified as markers of breast cancer. Twenty-three of the 166 genes (14%) were relatively tissue restricted, suggesting use as cancer-specific targets. We also identified several new markers of ovarian cancer. Our results indicate that CAST is a robust, rapid, and low cost method to identify cell surface and secreted proteins and is applicable to a variety of relevant biological questions. (Cancer Res 2005; 65(18): 8209-17)

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Section: -E Immunohistostaining Confirmed the Pcr Results And Showmentioning

confidence: 99%

Selective Identification of Secreted and Transmembrane Breast Cancer Markers using Escherichia coli Ampicillin Secretion Trap

Ferguson¹,

Muenster²,

Zang³

et al. 2005

Cancer Research

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“…Therefore, we did in vitro glycosylation assays to evaluate the capacity of this cell extract to glycosylate peptides from various human mucins. As shown in Table 1, we selected peptides derived from the tandem repeat of MUC1 (a mucin highly expressed in breast tissue whose O-glycosylated chains are shortened, or even absent, in breast cancer) and from other nonmammary mucins previously identified in breast cancer cell lines and breast carcinoma extracts, such as MUC2 (23), MUC5B (24), and MUC6 (25). All peptides were glycosylated, although at different rates (Fig.…”

Section: Glycosylation Of Mucin Peptides By Mcf-7 Cell Extractsmentioning

confidence: 99%

“…Identification of native MUC6 glycoproteins expressing the Tn antigen in MCF-7 cells. Previous works showed that MUC6 is expressed in breast cancer, including studies done on MCF-7 cells (25), pleural effusions associated to breast cancer (21), and mRNA expression of human breast tumors (24). To identify native MUC6 proteins bearing Tn antigen, we did Western blot analysis using the anti-Tn mAb 83D4 and two different anti-MUC6 antisera.…”

Section: Glycosylation Of Mucin Peptides By Mcf-7 Cell Extractsmentioning

confidence: 99%

“…An aberrant expression of MUC2 (23), MUC5B (24), and MUC6 (25) in breast cancer has been observed, suggesting that these apomucins are up-regulated during the course of malignant transformation of breast epithelium. It is possible that the aberrant expression of a specific type of apomucin could lead to the accumulation of simple O-glycan tumor-associated antigens, such as Tn.…”

Section: Cancer Researchmentioning

confidence: 99%

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Molecular Basis of Incomplete O-Glycan Synthesis in MCF-7 Breast Cancer Cells: Putative Role of MUC6 in Tn Antigen Expression

et al. 2005

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An incomplete elongation of O-glycan saccharide chains in mucins have been found in epithelial cancers, leading to the expression of shorter carbohydrate structures, such as the Tn antigen (GalNAc-O-Ser/Thr). This antigen is one of the most specific human cancer-associated structures and is capable of inducing effective immune responses against cancer cells. We aimed to investigate the causes of the expression of Tn antigen in the Tn-rich MCF-7 breast cancer cell line focusing on the first step of the O-glycosylation process. Interestingly, amino acid sequences derived from ''nonmammary'' apomucins (MUC5B and MUC6) were very good acceptor substrates for ppGalNAc-Ts, which are the enzymes catalyzing the Tn antigen synthesis. MUC6 peptide glycosylation with MCF-7 microsome extracts as source of ppGalNAc-T activity yielded 95% conversion of the peptide into MUC6-Tn. In addition, the MUC6-Tn glycopeptide was a poor acceptor substrate for core 1 B3Gal-T, the next enzyme involved in the saccharide chain biosynthesis, yielding only 5% conversion of MUC6-Tn into MUC6-TF. These results indicate that nonmammary apomucin expression could be responsible, at least in part, for Tn antigen expression in MCF-7 breast cancer cells due to a combined action on glycosyltransferases: an increase of ppGalNAc-T activity and a decrease of core 1 B3Gal-T activity.

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“…Our group has reported the expression of MUC5B in breast tissues, showing that MUC5B apomucin was detected in >80% of primary breast tumors while it was absent in normal control breast samples (27). Furthermore, the detection of MUC5B mRNA could constitute a specific marker applicable to the molecular diagnosis of breast cancer cell dissemination (30). Using different approaches we have demonstrated that the human breast cancer cell line MCF-7 expresses MUC5B (27).…”

Section: Introductionmentioning

confidence: 95%

MUC5B silencing reduces chemo-resistance of MCF-7 breast tumor cells and impairs maturation of dendritic cells

García¹,

Tiscornia

Libisch

et al. 2016

International Journal of Oncology

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Abstract. Mucins participate in cancer progression by regulating cell growth, adhesion, signaling, apoptosis or chemo-resistance to drugs. The secreted mucin MUC5B, the major component of the respiratory tract mucus, is aberrantly expressed in breast cancer, where it could constitute a cancer biomarker. In this study we evaluated the role of MUC5B in breast cancer by gene silencing the MUC5B expression with short hairpin RNA on MCF-7 cells. We found that MUC5B-silenced MCF-7 cells have a reduced capacity to grow, adhere and form cell colonies. Interestingly, MUC5B knock-down increased the sensitivity to death induced by chemotherapeutic drugs. We also show that MUC5B silencing impaired LPS-maturation of DCs, and production of cytokines. Furthermore, MUC5B knock-down also influenced DC-differentiation and activation since it resulted in an upregulation of IL-1β, IL-6 and IL-10, cytokines that might be involved in cancer progression. Thus, MUC5B could enhance the production of LPS-induced cytokines, suggesting that the use of MUC5B-based cancer vaccines combined with DC-maturation stimuli, could favor the induction of an antitumor immune response.

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Detection of bone marrow‐disseminated breast cancer cells using an RT‐PCR assay of MUC5B mRNA

Abstract: The evaluation of disseminated epithelial tumor cells in breast cancer patients has generated considerable interest due to its potential association with disease recurrence.

Cited by 31 publications

References 41 publications

Selective Identification of Secreted and Transmembrane Breast Cancer Markers using Escherichia coli Ampicillin Secretion Trap

Selective Identification of Secreted and Transmembrane Breast Cancer Markers using Escherichia coli Ampicillin Secretion Trap

Molecular Basis of Incomplete O-Glycan Synthesis in MCF-7 Breast Cancer Cells: Putative Role of MUC6 in Tn Antigen Expression

MUC5B silencing reduces chemo-resistance of MCF-7 breast tumor cells and impairs maturation of dendritic cells

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