Metalloproteinases in Endometrial Cancer—Are They Worth Measuring?

Endometrial cancer is one of the most common gynecological malignancies, yet the molecular mechanisms that lead to tumor development and progression are still not fully established. Matrix metalloproteinases (MMPs) are a group of enzymes that play an important role in carcinogenesis. They are proteases involved in the degradation of the extracellular matrix (ECM) that surrounds the tumor and the affected tissue allows cell detachment from the primary tumor causing local invasion and metastasis formation. Recent investigations demonstrate significantly increased metalloproteinase and metalloproteinase inhibitor levels in patients with endometrial cancer compared to those with normal endometrium. In this review, we aim to show their clinical significance and possible use in the diagnosis and treatment of patients with endometrial cancer. We have critically summarized and reviewed the research on the role of MMPs in endometrial cancer.

Dysregulation of Cytoskeleton Remodeling Drives Invasive Leading Cells Detachment

Detachment of cancer cells is the first step in tumor metastasis and malignancy. However, studies on the balance of initial tumor anchoring and detachment are limited. Herein, we revealed that the regulation of cytoskeleton proteins potentiates tumor detachment. The blockage of TGF-β1 using neutralizing antibodies induced cancer cell detachment in the Boyden chamber and 3D in-gel spheroid models. Moreover, treatment with latrunculin B, an actin polymerization inhibitor, enhanced cell dissociation by abolishing actin fibers, indicating that TGF-β1 mediates the formation of actin stress fibers, and is likely responsible for the dynamics of anchoring and detachment. Indeed, latrunculin B disrupted the formation of external TGF-β1-induced actin fibers and translocation of intracellular vinculin, a focal adhesion protein, resulting in the suppression of cell adhesion. Moreover, the silencing of vimentin substantially reduced cell adhesion and enhanced cell detachment, revealing that cell adhesion and focal adhesion protein translocation stimulated by TGF-β1 require vimentin. Using the 3D in-gel spheroid model, we found that latrunculin B suppressed the cell adhesion promoted by external TGF-β1, increasing the number of cells that penetrated the Matrigel and detached from the tumor spheres. Thus, cytoskeleton remodeling maintained the balance of cell anchoring and detachment, and the TGF-β1/vimentin/focal adhesion protein assembly axis was involved in the control dynamics of initial tumor detachment.

Evaluation of the cytotoxicity of an Ivorian aphrodisiac of natural origin

In Côte d'Ivoire, the market of aphrodisiacs of natural origin has been significantly growing over the last decades. The objective of this study was to evaluate the cytotoxicity of the most employed by Ivorian men, called Aphro. For that, Caco-2/TC7 enterocytes and Vero ATCC cells kidney epithelial cells grown in DMEM medium were exposed to Aphro at concentrations ranging from 216 to 864 µg/mL. Lactate dehydrogenase (LDH) released upon cell death was assayed after 1, 2 and 24h of exposure. In parallel, the effect of Apho was evaluated in differentiated confluent cells by measurements of the trans-epithelial electrical resistance. A time course and significant dose response related increase of LDH was measured in the culture medium of Caco-2/TC7 and Vero ATCC cells exposed to the aphrodisiac. The trans-epithelial electrical resistance of the Caco-2/TC7 cell monolayers also showed a significant decrease from 1400 Ω*cm2 at the onset of the experiment to 450 Ω*cm2 after 24 h of treatment. Cell detachment and structural disorganization were observed in both cell lines. This study reveals the strong cytotoxicity of Aphro extract on both cell lines.

A simple and intuitive methodology for estimating red blood cell surface antigen expression variation using optical cell-detachment technique

The analysis of surface antigens on cells, especially red blood cells (RBCs), has attracted increasing attention due to the recognition of antigenic variation that can facilitate early diagnoses. This paper presents an alternative methodology to estimate the variation of surface antigen expressions using an optical cell-detachment technique to validate the binding of individual RBCs stuck on corresponding antibody-coated surfaces. The detachment tests were implemented by an optical tweezers with gradually decreasing laser powers associated with serial antibody dilutions. Then, the antigen expression variation was estimated based on the known antibody dilution folds. The B- and B3-types of RBCs were selected for the demonstration subjects. With the semi-quantitative analysis, the proposed methodology was successfully verified for evaluating the variation of the RBC surface antigen expressions. The analysis result shows good consistency with the literature’s findings.

Inhibition of Matrix Metalloproteinases and Cancer Cell Detachment by Ru(II) Polypyridyl Complexes Containing 4,7-Diphenyl-1,10-phenanthroline Ligands—New Candidates for Antimetastatic Agents

Primary tumor targeting is the dominant approach in drug development, while metastasis is the leading cause of cancer death. Therefore, in addition to the cytotoxic activity of a series of Ru(II) polypyridyl complexes of the type [Ru(dip)2L]2+ (dip: 4,7-diphenyl-1,10-phenanthroline while L = dip; bpy: 2,2′-bipyridine; bpy-SC: bipyridine derivative bearing a semicarbazone 2-formylopyridine moiety; dpq, dpq(CH3)2, dpb: quinoxaline derivatives) their ability to inhibit cell detachment was investigated. In vitro studies performed on lung cancer A549 cells showed that they accumulate in cells very well and exhibit moderate cytotoxicity with IC50 ranging from 4 to 13 µM. Three of the studied compounds that have dip, bpy-SC, or dpb ligands after treatment of the cells with a non-toxic dose (<1/2IC50) enhanced their adhesion properties demonstrated by lower detachment in the trypsin resistance assay. The same complexes inhibited both MMP-2 and MMP-9 enzyme activities with IC50 ranging from 2 to 12 µM; however, the MMP-9 inhibition was stronger. More detailed studies for [Ru(dip)2(bpy-SC)]2+, which induced the greatest increase in cell adhesion, revealed that it is predominately accumulated in the cytoskeletal fraction of A549 cells. Moreover, cells treated with this compound showed the localization of MMP-9 to a greater extent also in the cytoskeleton. Taken together, our results indicate the possibility of a reduction of metastatic cells escaping from the primary lesion to the surrounding tissue by prevention of their detachment and by influencing the activity of MMP-2 and MMP-9.

Preparation of thermosensitive PNIPAm-based copolymer coated cytodex 3 microcarriers for efficient non-enzymatic cell harvesting during 3D culturing

Enzymatic detachment of cells might damage important features of cells and could affect subsequent function of cells in various applications. Therefore, non-enzymatic cell detachment using thermosensitive polymer matrix is necessary for maintaining cell quality after harvesting. In this study, we synthesized thermosensitive PNIPAm-co-AAc-b-PS and PNIPAm-co-AAm-b-PS copolymers and LCST was tuned near to body temperature. Then, polymer solutions (5% w/v, 10% w/v, and 20% w/v) were spin coated to prepare films for cell adhesion and thermal-induced cell detachment. The apha-step analysis and SEM image of the films suggested that the thickness of the films depends on the molecular weight and concentration which ranged from 206 nm to 1330 nm for PNIPAm-co-AAc-b-PS and 97.5 nm to 497 nm for PNIPAm-co-AAm-b-PS. The contact angles of the films verified that the polymer surface was moderately hydrophilic at 37°C. From cell attachment and detachment studies, RAW264.7 cells, were convincingly proliferated on the films to a confluent of >80 % within 48 days. However, relatively more cells were grown on PNIPAm-co-AAm-b-PS (5%w/v) films and thermal-induced cell detachment was more abundant in this formulation. As a result, commercial cytodex 3 microcarrier was coated with PNIPAm-co-AAm-b-PS (5%w/v) and interestingly enhanced cell detachment with preserved potential of recovery was observed at low temperature during 3D culturing. Thus, surface modification of microcarriers with PNIPAm-co-AAm-b-PS could be vital strategy for non-enzymatic cell dissociation and able to achieve adequate number of cells with maximum cell viability, and functionality for various cell-based applications. Keywords: surface coated microcarriers; thermosensitive polymer; non-enzymatic cell detachment

COMPUTATIONAL STUDY OF GEOMETRIC EFFECTS OF BOTTOM WALL MICROGROOVES ON CELL DOCKING INSIDE MICROFLUIDIC DEVICES

Cells docking inside microfluidic devices is effective in studying cell biology, cell-based biosensing, as well as drug screening. Furthermore, single cell and regularly cells docking inside the microstructure of microfluidic systems are advantageous in different analyses of single cells exposed to equal drug concentration and mechanical stimulus. In this study, we investigated bottom wall microgrooves with semicircular and rectangular geometries with different sizes which are suitable for single cell docking along the length of the microgroove in [Formula: see text]-direction and numerous cells docking regularly in one line inside the microgroove in a 3D microchannel. We used computational fluid dynamics to analyze the fluid recirculation area inside different microgrooves. The height of recirculation area in the bottom of microgroove could affect the cell’s attachment, and also materials delivery to attached cells, so the height of recirculation area may have optimum value. In addition, we analyzed the fluid drag force on cell movement toward the microgroove. This parameter was proportional to the fluid velocities in [Formula: see text] and [Formula: see text] directions in different microgrooves geometries. In different microgrooves’ geometries the fluid velocity in [Formula: see text]-direction did not change, but the fluid velocity in [Formula: see text]-direction decreased inside the microgroove. Therefore, the cell movement time inside the microgroove increased, and also the drag force in [Formula: see text]-direction could push the cells toward the bottom due to the lower drag force in [Formula: see text]-direction. The percentages of negative shear stress and average shear stress on the adhered cell surface were also calculated. The lower average shear stress, and negative shear stress around 50% on the cell surface were against cell detachment from the substrate. The results indicated that at the constant fluid inlet velocity and microchannel height, microgroove geometry and ratio of cell size to the microgroove size play pivotal roles in the cell initial adhesion to the substrate as well as the cell detachment.

Arthrospira platensis accelerates the formation of an endothelial cell monolayer and protects against endothelial cell detachment after bacterial contamination

Within the last years a comprehensive number of scientific studies demonstrated beneficial effect of Arthropira platensis (AP) as dietary supplement due to a high content of proteins, minerals and vitamins. Positive effects like promoting the immune system, reducing inflammation and an anti-oxidant capacity are reported. In this study, the effect of an aqueous AP extract on primary human venous endothelial cells (HUVEC) was investigated. In addition, the effect of AP on HUVEC treated with a bacterial toxin (lipopolysaccharide, LPA), inducing an activation of HUVEC and cellular detachment, was analyzed. Depending on the concentration of AP extract a significantly accelerated formation of an endothelial cell monolayer was observed. Furthermore, the detachment of HUVEC after LPA addition was dramatically reduced by AP. In conclusion, the data are promising and indicatory for an application of Arthrospira platensis in the clinical field.