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Role of Histone Methyltransferase in Breast Cancer
Published in Meenu Gupta, Rachna Jain, Arun Solanki, Fadi Al-Turjman, Cancer Prediction for Industrial IoT 4.0: A Machine Learning Perspective, 2021
Surekha Manhas, Zaved Ahmed Khan
The effectiveness of G9a inhibitors or DNMT inhibitors is easily recognizable in cellular-based activities for in in vitro studies. MTT assay has been used in different research approaches to evaluate drug or inhibitor effectiveness. In addition, the anti-tumor activity of different drugs is predictable by targeting the methyltransferase activity of G9a and DNMTs through MTS assay.
The Evolution of Anticancer Therapies
Published in David E. Thurston, Ilona Pysz, Chemistry and Pharmacology of Anticancer Drugs, 2021
Cell-based screens are also carried out by some companies, although this requires more complex equipment due to the need to control the temperature, gaseous environment, and sterility to ensure satisfactory cell growth. For example, tumor cells can be grown in multi-well plates, and individual compounds added to each well. A colorimetric detection method such as the MTT assay (Figure 2.7) can then be used to measure the number of dead versus living cells. This assay is based on the reduction of a yellow tetrazolium salt (MTT; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) by NAD(P)H-dependent oxidoreductase enzymes to purple formazan crystals by metabolically active cells. A. The cell-based MTT assay is based on conversion of the yellow MTT compound to the purple formazan via NAD(P)H-dependent oxidoreductase enzymes. B. A 96-well plate showing no effect on the cells in 92 of the wells, whereas the compounds in four wells on the left-hand side of the plate have killed the cells (i.e., the cells are not metabolizing the MTT).
Toxicity and Cellular Uptake of Gold Nanoparticles: What We Have Learned So Far *
Published in Valerio Voliani, Nanomaterials and Neoplasms, 2021
Alaaldin M. Alkilany, Catherine J. Murphy
There are many assays used to measure the cellular impact of a drug that can also be applied to measure the impact of nanoparticle exposure on cells. One common assay is the LDH assay, which is a colorimetric assay measuring the release of lactate dehydrogenase (LDH) into the culture media as an indicator of cellular membrane disruption (Marquis et al. 2009). A metabolic assay considered the “gold standard” for cytotoxicity is the MTT assay, which is a colorimetric assay that measures the enzymatic activity of cellular mitochondria. If cells properly metabolize the MTT dye, the cell culture will turn blue, allowing for simple absorbance measurements to be used to quantify cellular activity (Marquis et al. 2009).
Altered VDAC-HK association and apoptosis in mouse peripheral blood lymphocytes exposed to diabetic condition: an in vitro and in vivo study
Published in Archives of Physiology and Biochemistry, 2023
Melinda Nongbet Sohlang, Suktilang Majaw
MTT assay is a sensitive indicator of the metabolic viability indicative of mitochondrial function (Fiorillo et al. 2006). As shown in Figure 3(a), decreased mitochondrial activity (p < .001) was observed in PBL exposed to high Glc/PA as well as in that of diabetic PBL with p < .001. Changes in MMP directly contribute to mitochondria-mediated apoptosis. This change in MMP was analysed using JC-1 dye. Healthy cells with high MMP have JC-1 crossing the plasma membrane thereby forming aggregates that emit signal mostly at the red fluorescent channel while apoptotic cells have lower MMP where JC-1 remains in the cytoplasm as a monomeric form emitting a signal at the green channel. A lower MMP was observed in PBL treated with high Glc/PA and diabetic PBL (p < .001) compared to control groups (Figure 4(a)) as seen by the higher percentages of cells in the green fluorescence region (Figure 4(b)). This may therefore result in the release of apoptotic factors that initiate apoptosis by activating the death caspase indicated by caspase-3 (Porter and Janicke 1999).
Co-delivery of isotretinoin and clindamycin by phospholipid-based mixed micellar system confers synergistic effect for treatment of acne vulgaris
Published in Expert Opinion on Drug Delivery, 2021
Gajanand Sharma, Yukhti Yachha, Kanika Thakur, Akanksha Mahajan, Gurjeet Kaur, Bhupinder Singh, Kaisar Raza, OP Katare
Very few studies have reported the in vitro toxicity of topical retinoids like ITR on human skin cells. HaCat cells are the immortal human keratinocyte cells, which have been employed in many studies as standard for epidermal keratinocytes. Untreated HaCat cells served as the control. MTT assay is employed to measure the viable cells in 96-well plates without the need for cell counting; hence it is used to determine cytotoxicity of various drugs at different concentrations. The assay is based on the principle that mitochondrial activity remains almost constant for most of the viable cells. Therefore, the viable cell count is directly related to the mitochondrial activity. The results of cell viability assay using MTT dye are depicted in Figure 5. Control and the formulations (ITR-CLIN-loaded MM and Blank MM) did not show any toxicity till 100 µg/mL after 24 h of treatment indicating that all the excipients employed for the formulation of mixed micellar gel are safe to use without any cytotoxic effects. The differences in % cell viability between control and the developed formulations was non-significant; however, it was significant between control and pure drug (p < 0.05). The study concluded that the formulations are non-cytotoxic to HaCat cell lines [13,45].
Toll-like receptor (TLR) gene expression and immunostimulatory effect of CpG oligonucleotides in hormone receptor positive cell line T47D and triple negative breast cancer cell line MDA-MB-468
Published in Immunopharmacology and Immunotoxicology, 2020
Sudhakar Natarajan, Mohan Ranganathan
MTT assay is a colorimetric assay used to measure the cellular metabolic activity as an indicator of cellular proliferation, cell viability and cytotoxicity. The effect of CpG ODN on breast cancer cell lines T47D and MDA-MB-468 was evaluated by mitrochondrial-dependent reduction of (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) as described by Marshall et al [16]. For performing the MTT assay, 2 × 104 cells of T47D and MDA-MB-468 cells, respectively were seeded in a 96-well plate and incubated for 24 h. This was followed by stimulation of cells with CpG ODN 2006 in triplicate wells at a concentration of 1 µM, 2 µM and 3 µM for 24 h. Cells alone and media alone in quadruplicates were included in every experiment. After completion of 24 h, cell viability of CpG treated and untreated cells were checked by addition of 0.5 mg/ml of MTT (Sigma-aldrich); incubated for 4 h. Thereafter DMSO (Himedia, India) and isopropanol (Fisher Scientific) were used in 1:1 ratio to dissolve the formazan crystals. The absorbance was quantitated at 595 nm using iMark micro plate reader (Bio-Rad Laboratories, Inc.) Cell viability rate was determined using the following formula. Cell Viability rate (%) = (OD value of test/OD value of Control) X 100. The cell viability/cell cytotoxicity results were represented as mean ± sd of three independent experiments.