Methods to Study the Vasculature in ADPKD
Jinghua Hu, Yong Yu in Polycystic Kidney Disease, 2019
The wound-healing assay is a simple and economical method broadly used in many disciplines to study in vitro two-dimensional cell migration.138 The assay can be performed in standard well plates (from 12- to 96-well plates) by simply scratching a cell monolayer using a pipette tip as shown in Figure 7.6a.139,140 The cell-free gap induces directional cell migration, and by capturing images of migrating cells at fixed time intervals, the speed at which the wound is closed can be measured.141 The investigator can manipulate gene expression, extracellular matrix composition or other variables to investigate the mechanisms involved in cell migration. Appropriate controls should be performed and analyzed in parallel.142,143 A number of factors should be considered and controlled to maximize reproducibility, including consistently sizing the width/depth of the pipette wound and maintaining a constant cell density between experimental conditions and controls.144
Microalgae and Cyanobacteria as a Potential Source of Anticancer Compounds
Gokare A. Ravishankar, Ranga Rao Ambati in Handbook of Algal Technologies and Phytochemicals, 2019
There have been several reports on the anticancer effect of ATX against breast cancer based on both cell and animal models. For instance, McCall et al. (2018) demonstrated that treatment of ATX significantly reduced proliferation rates and inhibited breast cancer cell migration compared to the control normal breast epithelial cells. Inhibition of cancer cell migration is desirable as this would reduce the number of metastases formed. In addition, feeding of ATX was found to delay tumor growth and modulated immune response in a mouse cancer model (Nakao et al. 2010). The plasma levels of ATX as well as natural killer-cell subpopulation and plasma interferon-γ increased in mice that were fed ATX. However, the increase was only observed when ATX was given before tumor initiation, suggesting that an adequate blood ATX status is required to protect against tumor initiation. In another study, Yuri et al. (2016) compared the anticancer effects of ATX and canthaxanthin (CTX) in N-methyl-N-nitrosourea (MNU)-induced mammary cancer in a rat model. Feeding of ATX (0.4% diet) but not CTX reduced the incidence of palpable mammary carcinoma in the experimental animals. The study further showed that changes in adiponectin might be involved in the mechanism of action.
Identifying Nanotoxicity at the Cellular Level Using Electron Microscopy
Suresh C. Pillai, Yvonne Lang in Toxicity of Nanomaterials, 2019
ROS are generated by the incomplete reduction of oxygen within the cell. When maintained at adequate levels these molecules can interact with and contribute to cell migration signalling pathways and specifically with the actin cytoskeleton (Stanley et al., 2014). When homeostasis fails and management of levels of ROS within the cell exceed the beneficial level, oxidative stress leads to damage within the sample which may ultimately trigger apoptosis or cell death (Ray et al., 2012). Nanoparticles have been implicated in both ROS-dependent and -independent apoptotic pathways (Yang et al., 2014, Zhu et al., 2016). Previous work from our group, cited above as Stanley et al., has focused on the delicate interaction and balance of oxidation and oxidative stress within the cell. As the actin cytoskeleton, and its regulators the GTPase family, are so closely implicated with the endocytic mechanism, the effects of an up- or downregulation of ROS levels within the cell may also affect the endocytosis of nanoparticulate matter.
Expression and prognostic impact of NTF3 and TrkC in hepatocellular carcinoma
Published in Scandinavian Journal of Gastroenterology, 2023
Hejing Wang, Chenhan Zhong, Lina Qi, Xuefeng Fang, Ying Yuan
Cell migration was detected using wound healing and transwell assays. Regarding MHCC97-L cells, no significant difference was observed in the migration rate of the control and pCDH-NTF3 groups at 12 h (p = 0.530) and 24 h (p = 0.796) (Figure 3(C)). HepG2 cell mobility was similar in the control, pCDH-vector, and pCDH-NTF3 groups. At 12 h, the p-value of 0.144 illustrated the difference between the control and pCDH-NTF3 groups and the p-value 0.151 indicated the difference between the pCDH-vector and pCDH-NTF3 groups. At 24 h, no discrepancy was observed between the control and pCDH-NTF3 groups (p = 0.456). After 36 h, no discrepancy was observed in HepG2 cells between the pCDH-NTF3 and control (p = 0.359) and pCDH-vector (p = 0.603) groups. As shown in Figure3(D), after 48 h, no discrepancy was observed in MHCC97-L cells between the pCDH-NTF3 and control (p = 0.139) or pCDH-vector (p = 0.291) groups. Therefore, NTF3 overexpression had no significant effect on the migration ability of MHCC97-L and HepG2 cells.
Comparison of Virosome vs. Liposome as drug delivery vehicle using HepG2 and CaCo2 cell lines
Published in Journal of Microencapsulation, 2021
Varun Kumar, Ramesh Kumar, V. K. Jain, Suman Nagpal
A wound-healing assay was used to assess cell migration as previously described (Cory 2011). Briefly, 5 × 104 cells/plate were inoculated in 24-well plates for 24 h; and allowed to form a complete monolayer over the plate. The scratches were induced using a sterile pipette tip, the controlled wells (six) were remained untreated and 24 wells from group-IA & IIA and Group-IB & -IIB were treated with NC-virosome and NC-Liposome (25–100μg/ml), respectively. The Group-IC & -IIC were treated with Curcumin (25–100μg/ml). Images were captured at 0 h; and 48 h; to measure the wound width, the treated wells were compared with the untreated or controlled wells. The migration of cells was observed through inverted light microscopy. Wound closure can be calculated from the following formula (Yarrow et al. 2004, Rodriguez et al. 2005): t = Area of wound measured immediately after scratch
Antimetastatic Properties of Tea Polyphenols
Published in Nutrition and Cancer, 2020
Cell motility and migration is essential for organogenesis; however, when regulation fails, it may result in metastasis. Loss of epithelial characteristics (apical– basal polarity, differentiated, organized) and gaining mesenchymal-like cell phenotype are required for the acquisition of motility. It is suggested that the detachment and escaping of cells from the primary tumor mimics the developmental process known as epithelial–mesenchymal transition. An EMT is characterized by loss of cell-junction proteins, e.g., epithelial E-cadherin, an increased expression of mesenchymal markers, such as N-cadherin, vimentin intermediate filaments and fibronectin, loss of cell polarity, and gaining a spindle-shaped form (11,35). E- to N-cadherin expression is known as cadherin-switch that leads to enhanced motility (35). Besides increasing cell motility, EMT also helps to maintain a stem cell property, in suppressing apoptosis, senescence, immune reactions, and to acquire resistance to chemo- and radiotherapy (33). EMT in metastatic cells, however, is transitional (1). EMT is reversed and cells regain epithelial characteristics before they settle down at secondary sites. This phenomenon is known as mesenchymal–epithelial transition (MET). Invasion of metastatic cells occurs through the ECM by different mechanisms. They move either as single cell via mesenchymal cell migration or ameboid cell migration, or move collectively as epithelial sheet.