Time-Lapse 2D Imaging of Phagocytic Activity in M1 Macrophage-4T1 Mouse Mammary Carcinoma Cells in Co-cultures

Zaidi, Noorzaileen Eileena; Shazali, Nur Aima Hafiza; Leow, Adam Thean Chor; Osman, Mohamad; Ibrahim, Kamariah; Jaoi-Edward, Marilyn; Rahman, Nik Mohd Afizan Nik Abd

doi:10.3791/60281

Cited by 5 publications

(5 citation statements)

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“…This cell line has several qualities as a human mammary cancer model which is easily transplanted into the mammary gland , highly tumorigenic and invasive and easily metastasized to distant organs (28). In this experiment, RAW264.7 cells were polarized into M2 macrophages by the influences of M2-inducing cytokines interleukin-4 (IL-4) and interleukin-13 (IL-13) (22,23). RAW264.7 cells showed an altered morphology after being stimulated with IL4/ IL13 as illustrated by previous studies (22,23).…”

Section: Discussionmentioning

confidence: 60%

“…In this experiment, RAW264.7 cells were polarized into M2 macrophages by the influences of M2-inducing cytokines interleukin-4 (IL-4) and interleukin-13 (IL-13) (22,23). RAW264.7 cells showed an altered morphology after being stimulated with IL4/ IL13 as illustrated by previous studies (22,23). These polarized M2 macrophages also expressed the signature M2 macrophage marker, CD206 (29).…”

Section: Discussionmentioning

confidence: 61%

“…Alteration of the cell morphology was observed after the addition of (IL-4 and IL-13). The cells showed elongated, spindle-like shaped cells (22,23,24) (Fig. 1 A-B).…”

Section: Polarization Of Raw2647 Cells Into M2 Macrophagesmentioning

confidence: 99%

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Interleukin-27 Disrupts the Crosstalk of Apoptotic Activities between 4T1 Breast Cancer Cells and M2 Macrophages

Yusof¹,

Fuadi²,

Hamid³

et al. 2022

MJMHS

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Introduction: Cytokine immunotherapy such as Interleukin-27 (IL-27) has been foreseen as a promising alternative anti-cancer treatment. Thus, this study aimed to investigate whether IL-27 gene therapy regulates crosstalk between breast cancer cells and macrophages in the sense of pro-apoptotic activities. Methods: This study has led to the development of recombinant pcDNA3.4-IL27. The recombinant pcDNA3.4-IL27 was transfected into 4T1 murine mammary carcinoma cells alone and co-culture of 4T1 with M2 macrophages. The successful expression of IL-27 in the cells were determine through the immunofluorescence staining and detection of CD206, M2 macrophages marker. Apoptotic effects of pcDNA3.4-IL27 were assessed through MTT assay, Annexin V flow cytometer analysis, and AO/PI dual staining. Results: Our findings shows that pcDNA3.4-IL27 has the ability to induce apoptosis in both of the cell group and performs better in the co-culture of 4T1 with M2 macrophages compared to 4T1 cells alone. PcDNA3.4-IL27 induced apoptosis through the altered cell morphology and reduction in the number of viable cells. Conclusion: These data demonstrate that pcDNA3.4-IL27 has the ability to induce apoptosis in both 4T1 cell alone and co-cultured 4T1 with M2 macrophages. Thus, could serve as a potential anti cancer candidate against breast cancer.

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Section: Discussionmentioning

confidence: 60%

Section: Discussionmentioning

confidence: 61%

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Interleukin-27 Disrupts the Crosstalk of Apoptotic Activities between 4T1 Breast Cancer Cells and M2 Macrophages

Yusof¹,

Fuadi²,

Hamid³

et al. 2022

MJMHS

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“…Excess FAs taken up by CD36 are primarily converted to triacylglycerol (TAG) storage as cytoplasmic lipid droplets in adipose tissues while in skeletal muscles they are utilized in fatty acid oxidation (FAO) [ 26 , 27 ]. Macrophage ingests ox-LDL by binding and internalization in antigen-presenting cells [ 28 ], whereas dendritic cells phagocytose apoptotic cells both via the CD36 receptor [ 29 , 30 ]. Interestingly, CD36 also has been found to express on human skeletal muscle mitochondria membrane, where it is responsible for mitochondrial LCFAs transport and subsequent oxidation [ 31 ].…”

Section: Cd36 Receptor: Gene Structure Distribution and Functionmentioning

confidence: 99%

CD36-Fatty Acid-Mediated Metastasis via the Bidirectional Interactions of Cancer Cells and Macrophages

Zaidi

Shazali

Leow

et al. 2022

Cells

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Tumour heterogeneity refers to the complexity of cell subpopulations coexisting within the tumour microenvironment (TME), such as proliferating tumour cells, tumour stromal cells and infiltrating immune cells. The bidirectional interactions between cancer and the surrounding microenvironment mark the tumour survival and promotion functions, which allow the cancer cells to become invasive and initiate the metastatic cascade. Importantly, these interactions have been closely associated with metabolic reprogramming, which can modulate the differentiation and functions of immune cells and thus initiate the antitumour response. The purpose of this report is to review the CD36 receptor, a prominent cell receptor in metabolic activity specifically in fatty acid (FA) uptake, for the metabolic symbiosis of cancer–macrophage. In this review, we provide an update on metabolic communication between tumour cells and macrophages, as well as how the immunometabolism indirectly orchestrates the tumour metastasis.

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“…Differentiation of a monocyte precursor to an M2 macrophage phenotype is promoted by hypoxia and immunosuppressive cytokines, such as IL-4, IL-10 and IL-13 [ 37 , 38 ]. In contrast, antitumorigenic M1 macrophages facilitate tumor control via multiple mechanisms, including phagocytosis and secretion of proinflammatory cytokines, such as IFN-γ, IFN-β and IFN-α [ 39 ]. Recent efforts in single-cell RNA sequencing of solid tumors have identified multiple subclusters of TAMs, which suggests that macrophage functionality exists across a continuum of states, and therefore, binary classification inadequately represents complex TAM phenotypes.…”

Section: Key Cellular Components Of Innate Immunitymentioning

confidence: 99%

Targeting Innate Immunity in Cancer Therapy

et al. 2021

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The majority of current cancer immunotherapy strategies target and potentiate antitumor adaptive immune responses. Unfortunately, the efficacy of these treatments has been limited to a fraction of patients within a subset of tumor types, with an aggregate response rate of approximately 20% to date across all malignancies. The success of therapeutic inhibition of programmed death protein 1 (PD-1), protein death ligand 1 (PD-L1) and cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) with immune checkpoint inhibitors (ICI) has been limited to “hot” tumors characterized by preexisting T cell infiltration, whereas “cold” tumors, which lack T cell infiltration, have not achieved durable benefit. There are several mechanisms by which “cold” tumors fail to generate spontaneous immune infiltration, which converge upon the generation of an immunosuppressive tumor microenvironment (TME). The role of the innate immune system in tumor immunosurveillance and generation of antitumor immune responses has been long recognized. In recent years, novel strategies to target innate immunity in cancer therapy have emerged, including therapeutic stimulation of pattern recognition receptors (PRRs), such as Toll-like receptors (TLRs); the DNA sensing cGAS/STING pathway; nucleotide-binding oligomerization domain-like receptors (NLRs), such as NLRP3; and the retinoic acid-inducible gene-I (RIG-I)-like receptors (RLRs). In addition, therapeutic modulation of key innate immune cell types, such as macrophages and natural killer cells, has been investigated. Herein, we review therapeutic approaches to activate innate immunity within the TME to enhance antitumor immune responses, with the goal of disease eradication in “cold” tumors. In addition, we discuss rational immune-oncology combination strategies that activate both innate and adaptive immunity, with the potential to enhance the efficacy of current immunotherapeutic approaches.

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Time-Lapse 2D Imaging of Phagocytic Activity in M1 Macrophage-4T1 Mouse Mammary Carcinoma Cells in Co-cultures

Cited by 5 publications

References 0 publications

Interleukin-27 Disrupts the Crosstalk of Apoptotic Activities between 4T1 Breast Cancer Cells and M2 Macrophages

Interleukin-27 Disrupts the Crosstalk of Apoptotic Activities between 4T1 Breast Cancer Cells and M2 Macrophages

CD36-Fatty Acid-Mediated Metastasis via the Bidirectional Interactions of Cancer Cells and Macrophages

Targeting Innate Immunity in Cancer Therapy

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