Extracellular vesicles produced by mouse breast adenocarcinoma 4T1 cells with up- or down-regulation of adaptor protein Ruk/CIN85 differentially modulate the biological properties of 4T1 WT cells

Abstract BACKGROUND Extracellular vesicles (EVs) in biofluids are potential biomarkers of disease. To explore the clinical relevance of EVs, a specific generic EV marker would be useful, one that does not require antibodies and binds to all EVs. Here we evaluated 5 commonly used generic markers for flow cytometry. METHODS Flow cytometry (A60-Micro, Apogee) was used to evaluate the ability of the generic EV markers calcein acetoxymethyl ester, calcein acetoxymethyl ester violet, carboxyfluorescein succinimidyl ester (CFSE), 4-(2-[6-(dioctylamino)-2-naphthalenyl]ethenyl)-1-(3-sulfopropyl)pyridinium (di-8-ANEPPS), and lactadherin to stain EVs from MCF7 human breast adenocarcinoma cell line-conditioned culture medium [epithelial cell adhesion molecule positive (EpCAM+)] or platelet EVs from human plasma [integrin β3 positive (CD61+)]. Side scatter triggering was applied as a reference, and the influence of non-EV components (proteins and lipoproteins) was evaluated. RESULTS Di-8-ANEPPS, lactadherin, and side scatter detected 100% of EpCAM+ MCF7 EVs. Lactadherin and side scatter detected 33% and 61% of CD61+ EVs, respectively. Di-8-ANEPPS detected platelet EVs only if soluble protein was first removed. Because all generic markers stained proteins, at best 33% of platelet EVs in plasma were detected. The calcein markers and CFSE were either insensitive to EVs in both samples or associated with swarm detection. CONCLUSIONS None of the generic markers detected all and only EVs in plasma. Side scatter triggering detected the highest concentration of plasma EVs on our A60-Micro, followed by lactadherin. The choice between scatter or lactadherin primarily depends on the analytical sensitivity of the flow cytometer used.

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Co-localization of HIV-1 Nef with the AP-2 adaptor protein complex correlates with Nef-induced CD4 down-regulation

The EMBO Journal ◽

10.1093/emboj/16.23.6964 ◽

1997 ◽

Vol 16 (23) ◽

pp. 6964-6976 ◽

Cited By ~ 172

Author(s):

M. E. Greenberg

Keyword(s):

Protein Complex ◽

Adaptor Protein ◽

Down Regulation ◽

Hiv 1

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Abstract 2937: The regulation of FGFR signaling by RTK adaptor protein down-regulation through CROX (cluster regulation of RUNX) theory in DNPC

10.1158/1538-7445.am2020-2937 ◽

2020 ◽

Author(s):

Asami Sasaski ◽

Youhei Yanagida ◽

Hiroshi Sugiyama ◽

Souichi Adachhi ◽

Yasuhiko Kamikubo

Keyword(s):

Adaptor Protein ◽

Down Regulation ◽

Fgfr Signaling

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Poly Organotin Acetates against DNA with Possible Implementation on Human Breast Cancer

International Journal of Molecular Sciences ◽

10.3390/ijms19072055 ◽

2018 ◽

Vol 19 (7) ◽

pp. 2055 ◽

Cited By ~ 11

Author(s):

George Latsis ◽

Christina Banti ◽

Nikolaos Kourkoumelis ◽

Constantina Papatriantafyllopoulou ◽

Nikos Panagiotou ◽

...

Keyword(s):

Binding Constants ◽

Fetal Lung ◽

Biological Properties ◽

Docking Studies ◽

Molecular Structures ◽

Breast Adenocarcinoma ◽

Human Breast ◽

X Ray Diffraction ◽

Ct Dna

Two known tin-based polymers of formula {[R3Sn(CH3COO)]n} where R = n-Bu– (1) and R = Ph– (2),were evaluated for their in vitro biological properties. The compounds were characterized via their physical properties and FT-IR, 119Sn Mössbauer, and 1H NMR spectroscopic data. The molecular structures were confirmed by single-crystal X-Ray diffraction crystallography. The geometry around the tin(IV) ion is trigonal bi-pyramidal. Variations in O–Sn–O···Sn′ torsion angles lead to zig-zag and helical supramolecular assemblies for 1 and 2, respectively. The in vitro cell viability against human breast adenocarcinoma cancer cell lines: MCF-7 positive to estrogens receptors (ERs) and MDA-MB-231 negative to ERs upon their incubation with 1 and 2 was investigated. Their toxicity has been studied against normal human fetal lung fibroblast cells (MRC-5). Compounds 1 and 2 exhibit 134 and 223-fold respectively stronger antiproliferative activity against MDA-MB-231 than cisplatin. The type of the cell death caused by 1 or 2 was also determined using flow cytometry assay. The binding affinity of 1 and 2 towards the CT-DNA was suspected from the differentiation of the viscosity which occurred in the solution containing increasing amounts of 1 and 2. Changes in fluorescent emission light of Ethidium bromide (EB) in the presence of DNA confirmed the intercalation mode of interactions into DNA of both complexes 1 and 2 which have been ascertained from viscosity measurements. The corresponding apparent binding constants (Kapp) of 1 and 2 towards CT-DNA calculated through fluorescence spectra are 4.9 × 104 (1) and 7.3 × 104 (2) M−1 respectively. Finally, the type of DNA binding interactions with 1 and 2 was confirmed by docking studies.

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Nanocapsules for the co-delivery of selol and doxorubicin to breast adenocarcinoma 4T1 cells in vitro

Artificial Cells Nanomedicine and Biotechnology ◽

10.1080/21691401.2017.1408020 ◽

2017 ◽

pp. 1-11 ◽

Cited By ~ 4

Author(s):

Rayane Ganassin ◽

Carolin Merker ◽

Mosar Corrêa Rodrigues ◽

Nayara Felipe Guimarães ◽

Carine Sampaio Cerqueira Sodré ◽

...

Keyword(s):

Breast Adenocarcinoma ◽

4T1 Cells

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Down-regulation of the Tumor Suppressor C-terminal Src Kinase (Csk)-binding Protein (Cbp)/PAG1 Is Mediated by Epigenetic Histone Modifications via the Mitogen-activated Protein Kinase (MAPK)/Phosphatidylinositol 3-Kinase (PI3K) Pathway

Journal of Biological Chemistry ◽

10.1074/jbc.m110.195362 ◽

2011 ◽

Vol 286 (18) ◽

pp. 15698-15706 ◽

Cited By ~ 25

Author(s):

Kei Suzuki ◽

Chitose Oneyama ◽

Hironobu Kimura ◽

Shoji Tajima ◽

Masato Okada

Keyword(s):

Tumor Suppressor ◽

Cancer Cells ◽

Histone Modifications ◽

Human Cancer ◽

Methylation Status ◽

Adaptor Protein ◽

Pi3k Pathway ◽

Mitogen Activated Protein Kinase ◽

Down Regulation ◽

Human Cancer Cells

The transmembrane adaptor protein Cbp (or PAG1) functions as a suppressor of Src-mediated tumor progression by promoting the inactivation of Src. The expression of Cbp is down-regulated in Src-transformed cells and in various human cancer cells, suggesting a potential role for Cbp as a tumor suppressor. However, the mechanisms underlying the down-regulation of Cbp remain unknown. The present study shows that Cbp expression is down-regulated by epigenetic histone modifications via the MAPK/PI3K pathway. In mouse embryonic fibroblasts, transformation by oncogenic Src and Ras induced a marked down-regulation of Cbp expression. The levels of Cbp expression were inversely correlated with the activity of MEK and Akt, and Cbp down-regulation was suppressed by inhibiting MEK and PI3K. Src transformation did not affect the stability of Cbp mRNA, the transcriptional activity of the cbp promoter, or the DNA methylation status of the cbp promoter CpG islands. However, Cbp expression was restored by treatment with histone deacetylase (HDAC) inhibitors and by siRNA-mediated knockdown of HDAC1/2. Src transformation significantly decreased the acetylation levels of histone H4 and increased the trimethylation levels of histone H3 lysine 27 in the cbp promoter. EGF-induced Cbp down-regulation was also suppressed by inhibiting MEK and HDAC. Furthermore, the inhibition of MEK or HDAC restored Cbp expression in human cancer cells harboring Cbp down-regulation through promoter hypomethylation. These findings suggest that Cbp down-regulation is primarily mediated by epigenetic histone modifications via oncogenic MAPK/PI3K pathways in a subset of cancer cells.

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Down-Regulation of Toll-Like Receptor 5 (TLR5) Increased VEGFR Expression in Triple Negative Breast Cancer (TNBC) Based on Radionuclide Imaging

Frontiers in Oncology ◽

10.3389/fonc.2021.708047 ◽

2021 ◽

Vol 11 ◽

Author(s):

Wen Jiang ◽

Yeming Han ◽

Ting Liang ◽

Chao Zhang ◽

Feng Gao ◽

...

Keyword(s):

Triple Negative ◽

Whole Body ◽

Tumor Uptake ◽

Toll Like Receptor ◽

Rt Pcr ◽

Down Regulation ◽

Tumor Bearing ◽

4T1 Cells ◽

Toll Like Receptor 5 ◽

Tlr5 Expression

In this study, GFP-tagged TNBC 4T1 cells with down-regulated TLR5 expression (TLR5− 4T1) and normal TLR5 expression (TLR5+ 4T1) were constructed, respectively. RT-PCR and Western blot studies showed that down-regulation of TLR5 obviously increased the expression of VEGFR in 4T1 cells. Highly stable radio-probes 125I-anti-TLR5 mAb/125I-VEGF/125I-IgG were obtained with labeling rates over 85% and radiochemical purities above 90%. Among these three probes, 125I−anti−TLR5 mAb and 125I-VEGF were used for specifically imaging TNBC, while 125I-IgG was used for comparison. Whole-body phosphorus autoradiography showed clear imaging at 48 h after injection of 125I-anti-TLR5 mAb and 125I-VEGF also provided clear imaging at 24 h. Biodistribution study demonstrated a higher tumor uptake of 125I-anti-TLR5 mAb in TLR5+ group compared with that in TLR5− group (P < 0.05), whereas tumor uptake of 125I-VEGF in TLR5+ group was lower than that in the TLR5− group (P < 0.05). Immunohistochemical staining suggested that the expression of TLR5 was lower, whereas the expression of VEGFR, CD31, and MVD (microvessel density) was higher in TLR5− tumor-bearing mice. In summary, the down-regulation of TLR5 in TNBC promoted the VEGFR expression and angiogenesis, resulting in the proliferation of TNBC cells. TLR5/VEGF might be a better indicator for monitoring the development of TNBC.

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