NCAM and attached polysialic acid affect behaviors of breast epithelial cells through differential signaling pathways

Abstract Background Neural cell adhesion molecule (NCAM), a common mammalian cell surface glycoprotein, is the major substrate of polysialic acid (PSA). Polysialylated NCAM occurs in many types of cancer, but rarely in normal adult tissues. Epithelial-mesenchymal transition (EMT) is an important process contributing to tumor metastasis. The functional role of NCAM hypersialylation in EMT is unclear. Method Expression of NCAM and PSA-NCAM in breast cancer progression were evaluated by western blot (WB), immunohistochemistry, semi-quantitative PCR and immunoprecipitation (IP). Overexpression of NCAM-140 and STX were performed to assess the functional role of NCAM hypersialylation by liposome transfection. Then, the cell proliferation ability was investigated with MTT assay. Transwell and wound closure assay were conducted to evaluate cell migratory ability. Phagokinetic gold sol assay and cell adhesion assay were performed to assess cell motility ability and cell adhesion ability, respectively. Furthermore, WB and IP were used to reveal the activated signaling pathway. Results The present study indicates that NCAM and attached polysialic acid affect behaviors of breast epithelial cells through differential signaling pathways, based on the following observations: (i) NCAM and PSA-NCAM were aberrantly regulated in breast cancer cells; (ii) NCAM and PSA-NCAM expression were upregulated in normal breast epithelial cells undergoing EMT; (iii) NCAM overexpression induced EMT in breast epithelial cells; (iv) NCAM promoted cell proliferation and migration through activation of a β-catenin/slug signaling pathway; (v) modification of PSA attached to NCAM inhibited cell adhesion and promoted cell motility through activation of an EGFR/STAT3 pathway. Conclusion These observations help clarify the molecular mechanisms whereby PSA and its major substrate NCAM modulate cell behaviors, and the significance of increased PSA expression on NCAM during EMT and tumor development.

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NCAM and attached polysialic acid affect behaviors of breast epithelial cells through differential signaling pathways

10.21203/rs.2.24259/v2 ◽

2020 ◽

Author(s):

Lin Cao ◽

Xin Wang ◽

Juhong Yang ◽

Jia Guo ◽

Xiang Li ◽

...

Keyword(s):

Breast Cancer ◽

Cell Adhesion ◽

Epithelial Cells ◽

Signaling Pathway ◽

Functional Role ◽

Polysialic Acid ◽

Breast Epithelial Cells ◽

Differential Signaling ◽

Breast Epithelial

Abstract Background Neural cell adhesion molecule (NCAM), a common mammalian cell surface glycoprotein, is the major substrate of polysialic acid (polySia). Polysialylated NCAM occurs in many types of cancer, but rarely in normal adult tissues. Epithelial-mesenchymal transition (EMT) is an important process contributing to tumor metastasis. The functional role of NCAM hypersialylation in EMT is unclear. Method Expression of NCAM and polysialylated NCAM in breast cancer progression were evaluated by western blot (WB), immunohistochemistry, semi-quantitative PCR and immunoprecipitation (IP). Overexpression of NCAM-140 and ST8SiaII were performed to assess the functional role of NCAM hypersialylation by liposome transfection. Cell proliferation ability was investigated with MTT assay. Transwell and wound closure assay were conducted to evaluate cell migratory ability. Phagokinetic gold sol assay and cell adhesion assay were performed to assess cell motility ability and cell adhesion ability, respectively. Furthermore, WB and IP were used to reveal the activated signaling pathway. Results The present study indicates that NCAM and attached polysialic acid affect behaviors of breast epithelial cells through differential signaling pathways, based on the following observations: (i) NCAM and polysialylated NCAM were aberrantly regulated in breast cancer cells; (ii) NCAM and polysialylated NCAM expression were upregulated in normal breast epithelial cells undergoing EMT; (iii) NCAM overexpression induced EMT in breast epithelial cells; (iv) NCAM promoted cell proliferation and migration through activation of a β-catenin/slug signaling pathway; (v) modification of polySia attached to NCAM inhibited cell adhesion and promoted cell motility through activation of an EGFR/STAT3 pathway. Conclusion This study demonstrates that NCAM and polysialylated NCAM facilitate different signaling pathway and affect different cell behaviors. Switching between the NCAM-mediated pathways appeared to depend on polySia decoration.

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NCAM and attached polysialic acid affect behaviors of breast epithelial cells through differential signaling pathways

10.21203/rs.2.24259/v3 ◽

2020 ◽

Author(s):

Lin Cao ◽

Xin Wang ◽

Juhong Yang ◽

Jia Guo ◽

Xiang Li ◽

...

Keyword(s):

Breast Cancer ◽

Cell Adhesion ◽

Epithelial Cells ◽

Signaling Pathway ◽

Functional Role ◽

Polysialic Acid ◽

Breast Epithelial Cells ◽

Differential Signaling ◽

Breast Epithelial

Abstract Background Neural cell adhesion molecule (NCAM), a common mammalian cell surface glycoprotein, is the major substrate of polysialic acid (polySia). Polysialylated NCAM occurs in many types of cancer, but rarely in normal adult tissues. Epithelial-mesenchymal transition (EMT) is an important process contributing to tumor metastasis. The functional role of NCAM hypersialylation in EMT is unclear. Method Expression of NCAM and polysialylated NCAM in breast cancer progression were evaluated by western blot (WB), immunohistochemistry, semi-quantitative PCR and immunoprecipitation (IP). Overexpression of NCAM-140 and ST8SiaII were performed to assess the functional role of NCAM hypersialylation by liposome transfection. Cell proliferation ability was investigated with MTT assay. Transwell and wound closure assay were conducted to evaluate cell migratory ability. Phagokinetic gold sol assay and cell adhesion assay were performed to assess cell motility ability and cell adhesion ability, respectively. Furthermore, WB and IP were used to reveal the activated signaling pathway. Results The present study indicates that NCAM and attached polysialic acid affect behaviors of breast epithelial cells through differential signaling pathways, based on the following observations: (i) NCAM and polysialylated NCAM were aberrantly regulated in breast cancer cells; (ii) NCAM and polysialylated NCAM expression were upregulated in normal breast epithelial cells undergoing EMT; (iii) NCAM overexpression induced EMT in breast epithelial cells; (iv) NCAM promoted cell proliferation and migration through activation of a β-catenin/slug signaling pathway; (v) modification of polySia attached to NCAM inhibited cell adhesion and promoted cell motility through activation of an EGFR/STAT3 pathway.Conclusion This study demonstrates that NCAM and polysialylated NCAM facilitate different signaling pathway and affect different cell behaviors. Switching between the NCAM-mediated pathways appeared to depend on polySia decoration.

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Role of Sphk1 in the malignant transformation of breast epithelial cells and breast cancer progression

Indian Journal of Cancer ◽

10.4103/0019-509x.175343 ◽

2014 ◽

Vol 51 (4) ◽

pp. 524 ◽

Cited By ~ 4

Author(s):

Y Zhang ◽

J Jiang ◽

XD Zheng ◽

Y Zhang ◽

XW Qi ◽

...

Keyword(s):

Breast Cancer ◽

Epithelial Cells ◽

Malignant Transformation ◽

Cancer Progression ◽

Breast Cancer Progression ◽

Breast Epithelial Cells ◽

Breast Epithelial

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Role of the Microenvironment in Regulating Normal and Cancer Stem Cell Activity: Implications for Breast Cancer Progression and Therapy Response

Cancers ◽

10.3390/cancers11091240 ◽

2019 ◽

Vol 11 (9) ◽

pp. 1240 ◽

Cited By ~ 10

Author(s):

Vasudeva Bhat ◽

Alison L. Allan ◽

Afshin Raouf

Keyword(s):

Breast Cancer ◽

Stem Cells ◽

Epithelial Cells ◽

Cancer Stem Cells ◽

Breast Tissue ◽

Therapy Response ◽

Breast Epithelial Cells ◽

Tissue Microenvironment ◽

Breast Epithelial

The epithelial cells in an adult woman’s breast tissue are continuously replaced throughout their reproductive life during pregnancy and estrus cycles. Such extensive epithelial cell turnover is governed by the primitive mammary stem cells (MaSCs) that proliferate and differentiate into bipotential and lineage-restricted progenitors that ultimately generate the mature breast epithelial cells. These cellular processes are orchestrated by tightly-regulated paracrine signals and crosstalk between breast epithelial cells and their tissue microenvironment. However, current evidence suggests that alterations to the communication between MaSCs, epithelial progenitors and their microenvironment plays an important role in breast carcinogenesis. In this article, we review the current knowledge regarding the role of the breast tissue microenvironment in regulating the special functions of normal and cancer stem cells. Understanding the crosstalk between MaSCs and their microenvironment will provide new insights into how an altered breast tissue microenvironment could contribute to breast cancer development, progression and therapy response and the implications of this for the development of novel therapeutic strategies to target cancer stem cells.

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Endotoxin Triggers Tumor Initiation Events in Nontumorigenic Breast Epithelial Cells and Enhances Invasion-Related Phenotype in Pretumorigenic and Tumorigenic Breast Epithelial Cells

International Journal of Inflammation ◽

10.1155/2021/4666380 ◽

2021 ◽

Vol 2021 ◽

pp. 1-13

Author(s):

Farah Yassine ◽

Sabreen F. Fostok ◽

Nataly Naser Al Deen ◽

Rabih S. Talhouk

Keyword(s):

Breast Cancer ◽

Epithelial Cells ◽

Human Breast ◽

Short Term ◽

Breast Epithelial Cells ◽

Inflammatory Phenotype ◽

Breast Cells ◽

Breast Epithelial

Inflammation is associated with the development of several cancers, including breast cancer. However, the molecular mechanisms driving breast cancer initiation or enhancement by inflammation are yet to be deciphered. Hence, we opted to investigate the role of inflammation in initiating and enhancing tumor-like phenotypes in nontumorigenic, pretumorigenic, and tumorigenic breast epithelial cells. Noncytotoxic endotoxin (ET) concentrations capable of inducing an inflammatory phenotype were determined for the different cell lines. Results showed that short-term ET exposure upregulated matrix metalloproteinase-9 (MMP-9) activity in nontumorigenic mammary epithelial cells of mouse (SCp2) and human origins (HMT-3522 S1; S1) and upregulated inflammatory mediators including nitric oxide (NO) and interleukin 1-β in tumorigenic human breast cells (MDA-MB-231), all in a dose-dependent manner. Long-term ET treatment, but not short-term, triggered the migration of SCp2 cells, and proliferation and migration of tumorigenic human breast cells MCF-7 and MDA-MB-231. Both short- and long-term ET exposures preferentially enhanced the invasion of pretumorigenic S1-connexin 43 knockout (Cx43-KO S1) cells compared to their nontumorigenic S1 counterparts. Moreover, both ET exposures disrupted lumen formation and apicolateral distribution of β-catenin in 3D cultures of S1 cells. In conclusion, ET treatment at concentrations that elicited inflammatory phenotype triggered tumor initiation events in nontumorigenic and pretumorigenic breast cells, and increased tumorigenicity of breast cancer cells. Our findings highlight the role of inflammation in enhancing migration, invasion, and loss of normal 3D morphology and suggest that such inflammatory insults can “add injury” to pretumorigenic and tumorigenic breast epithelial cells.

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Role of c-Myc in Estrogen Stimulated Cell Proliferation in Normal and Malignant Breast Epithelial Cells

10.21236/ada398172 ◽

2001 ◽

Author(s):

Eileen Rogen

Keyword(s):

Cell Proliferation ◽

Epithelial Cells ◽

Breast Epithelial Cells ◽

Malignant Breast ◽

Breast Epithelial

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Inhibition of MCF-7 breast cancer cell proliferation by MCF-10A breast epithelial cells in coculture

Cell Biology International ◽

10.1016/j.cellbi.2005.11.006 ◽

2006 ◽

Vol 30 (3) ◽

pp. 227-238 ◽

Cited By ~ 25

Author(s):

Barbara C. Spink ◽

Richard W. Cole ◽

Barbara H. Katz ◽

John F. Gierthy ◽

Laurie M. Bradley ◽

...

Keyword(s):

Breast Cancer ◽

Cell Proliferation ◽

Epithelial Cells ◽

Cancer Cell ◽

Breast Cancer Cell ◽

Cancer Cell Proliferation ◽

Breast Cancer Cell Proliferation ◽

Breast Epithelial Cells ◽

Breast Epithelial ◽

Mcf 7

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The Role of Redox Proteins in Arresting Proliferation of Breast Epithelial Cells Under Oxidative Stress

Annals of the Russian academy of medical sciences ◽

10.15690/vramn1030 ◽

2018 ◽

Vol 73 (5) ◽

pp. 289-293

Author(s):

Evgeniya V. Shakhristova ◽

Elena A. Stepovaya ◽

Evgeniy V. Rudikov ◽

Olga S. Sushitskaya ◽

Daria O. Rodionova ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Cycle ◽

Cell Proliferation ◽

Epithelial Cells ◽

Breast Epithelial Cells ◽

Redox Proteins ◽

Cell Cycle Inhibitor ◽

A Cell ◽

Breast Epithelial

Background: Redox status imbalance against the backdrop of oxidative stress development underlies the pathogenesis of a whole range of diseases. Many intracellular proteins contain free thiol groups and undergo redox regulation which is one of the key processes in controlling cell proliferation. Thioredoxin and glutaredoxin are involved in maintaining intracellular redox homeostasis and act as candidates in regulating proliferation. This provides prospects for future development of methods for diagnosis and targeted therapy of socially sensitive diseases accompanied by oxidative stress. The aim of the study is to reveal the role of redox proteins in molecular mechanisms of regulating HBL-100 breast epithelial cell proliferation under the effect of roscovitine, a cell cycle inhibitor. Materials and methods: Two research groups were formed. They included HBL-100 human breast epithelial cells incubated in the presence and absence of 20 mcM roscovitine for 18 hours. The intracellular thioredoxin levels were determined using Western blot analysis with specific monoclonal antibodies. Distribution of the cells among cell cycle phases were evaluated by flow cytometry. The activity of glutathione reductase, glutathione peroxidase, and thioredoxin reductase were measured by spectrophotometry. Results: Under the effect of roscovitine in the HBL-100 cells, cell cycle arrest in the G2/М phases occurred and oxidative stress developed. In the meantime, the decrease in the thioredoxin and glutaredoxin concentrations was registered along with the change in the functional activity of glutathione-dependent enzymes. Conclusions: Application of roscovitine, a cell cycle inhibitor, allowed creating a model of oxidative stress in the breast epithelial cells against the backdrop of inhibited cell proliferation. We identified that thioredoxin and glutaredoxin contributed to impairment of cell cycle progression. It points at a possibility to regulate cell proliferation by modulating the functional features of cellular redox-dependent proteins in different pathologies accompanied by oxidative stress.

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