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In-vitro Anti-Proliferative Screening in Selected Cassia Species
Published in Brijesh Kumar, Vikas Bajpai, Vikaskumar Gond, Subhashis Pal, Naibedya Chattopadhyay, Phytochemistry of Plants of Genus Cassia, 2021
Brijesh Kumar, Vikas Bajpai, Vikaskumar Gond, Subhashis Pal, Naibedya Chattopadhyay
A colorimetric sulforhodamine B (SRB) assay is the most preferred technique and was used for the measurement of anti-proliferative activity (Fricker and Buckley, 1995; Keepers et al., 1991; Skehan et al., 1990; Adaramoye et al., 2011). This basically depends on the incur of the negatively charged pink amino xanthine dye, SRB through basic amino acids in the cells. The dye released will give more acute colour and more absorbance, when the number of cells and amount of dye is taken up is greater, after fixing, the cells are lysed (Skehan et al., 1990). The SRB assay is sensitive, reproducible and gives better linearity, a good signal-to-noise ratio and has a stable end-point that does not require a time-sensitive measurement (Fricker and Buckley, 1995; Keepers et al., 1991). Ten thousand cells were seeded to each well of 96-well plate, grown overnight and exposed to test samples at 100 µg/ml concentration for 48 h. Cells were then fixed with ice-cold trichloroacetic acid (50% w/v, 50 µl/well), stained with SRB (0.4% w/v in 1% acetic acid, 50µl/well), washed and air dried. Bound dye was dissolved in 150 μL of 10 mM Tris base and plates were read at 510 nm absorbance (Epoch Microplate Reader, Biotek, USA).
Quantum Dots as Biointeractive and Non-Agglomerated Nanoscale Fillers for Dental Resins
Published in Mary Anne S. Melo, Designing Bioactive Polymeric Materials for Restorative Dentistry, 2020
Isadora Martini Garcia, Fabrício Mezzomo Collares
For the cytotoxicity evaluation, the polymerized samples were stored for 24 h in a culture medium. Then, this medium with possible leaching (eluate) from the samples was placed on human pulp cells. Cells and eluates were kept in contact for 72 h. The viability of cells after this period was analyzed via sulforhodamine B (SRB) colorimetric assay, which determines the cell density by staining proteins of viable cells. The viability of cells, according to ISO 10993-5 standardization, must be at least 70% to infer that the material has no cytotoxicity. The adhesives with and without ZnOQDs showed values higher than 70%, without difference between them. The method used in this study is an exciting alternative to the typical assay using tetrazolium salt (MTT). SRB method has the advantage not to be related to metabolic activity, but to enable cell enumeration based on protein content. Fewer compounds (intracellular and extracellular) interfere in the SRB outcome in comparison to MTT assay, and SRB shows a better predictive power than MTT (van Tonder et al. 2015).
Choerospondias axillaris (Hog plum)
Published in Mahendra Rai, Shandesh Bhattarai, Chistiane M. Feitosa, Wild Plants, 2020
Flow cytometry and sulforhodamine B (SRB) methods were employed to evaluate the antitumor activity of the compounds. In SRB assay, the compounds inhibited the proliferation of human cancer HCT-15 and HeLa cells. Flow cytometric analysis indicated that compounds pinocembrin and naringenin slightly inhibited the cell cycle of tsFT210 cells at the G2/M phase at higher concentrations, while dibutyl phthalate showed strong cytotoxicity at a higher concentration, but at a lower concentration, inhibited the cell cycle at the G0/G1 phase (Li et al. 2005).
Anticancer potentials of metformin loaded coconut oil nanoemulsion on MCF-7, HepG2 and HCT-116 cell lines
Published in Artificial Cells, Nanomedicine, and Biotechnology, 2023
Hadil Faris Alotaibi, El-Sayed Khafagy, Amr Selim Abu Lila, Haifa F. Alotaibe, Serag Eldin Elbehairi, Ashwag S. Alanazi, Mohammad Y. Alfaifi, Jawaher Abdullah Alamoudi, Sarah Salem Alamrani, Fatma Alzahraa Mokhtar
The sulforhodamine B (SRB) assay is one of the most commonly used colorimetric assays for evaluating cell viability via measuring cellular protein content. Consequently, an SRB assay was adopted to assess the cytotoxic potential of metformin-loaded coconut oil nanoemulsion against MCF-7, HepG2, and HCT 116 cells. As depicted in Figures 3–5, metformin remarkably lowered the viability of all tested cell lines in a dose-dependent manner. The IC50 values observed on various cell lines are summarized in Table 3. It was evident that metformin has a potent potential activity towards MCF-7 cancer cells, with IC50 19.61 ± 1.3 µg/mL and promising effect on HepG2 and HCT 116 cells with IC50 values of 18.17 ± 1.33 and 6.26 ± 02.1 µg/mL, respectively. Plain coconut oil nanoemulsion exerted a pronounced cytotoxic effect against all the tested cell lines, with a lower cytotoxic effect against MCF7 cells (IC50 44.72 ± 4.3 µg/mL) compared to either HepG2 or HCT116 cancer cells (IC50 values were 24.84 ± 3.9 µg/mL and 35.5 ± 4.2 µg/mL, respectively.
In silico identification and biological evaluation of a selective MAP4K4 inhibitor against pancreatic cancer
Published in Journal of Enzyme Inhibition and Medicinal Chemistry, 2023
Chao-Di Chang, Min-Wu Chao, Hsueh-Yun Lee, Yi-Ting Liu, Huang-Ju Tu, Ssu-Ting Lien, Tony Eight Lin, Tzu-Ying Sung, Shih-Chung Yen, Sing-Han Huang, Kai-Cheng Hsu, Shiow-Lin Pan
A sulforhodamine B (SRB) assay was used to measure the antiproliferative effect of a compound. The SRB assay was performed using a standard method as described previously39. Two pancreatic cancer cell lines were seeded in 96-well plates at a density of 5,000 cells per well overnight. The next day, 10% of trichloroacetic acid (TCA) was added to the basal group to maintain the cells at the initial density. After treating with different concentrations of a compound dissolved in 0.1% dimethyl sulfoxide (DMSO) for 72 and 96 h, cells were fixed with TCA for an additional 15 min and stained with 0.4% SRB in 1% acetic acid for 5 min. Then, the cells were washed three times with 1% acetic acid after 5–10 min of SRB staining to wash out unbound SRB crystals. The cells were dissolved in a 10 mM Tris base solution. At last, the absorbance at 515 nm was measured using a microplate reader. The absorbance of each well was normalised with the basal group, which was treated with DMSO. The concentration that led to 50% cell growth inhibition was determined using non-linear regression analysis (GI50, 50% growth inhibition).
Unusual dimeric flavonoids (brachydins) induce ultrastructural membrane alterations associated with antitumor activity in cancer cell lines
Published in Drug and Chemical Toxicology, 2023
Vera Lucia Maciel-Silva, Claudia Quintino da Rocha, Luciana Magalhães Rebelo Alencar, Patrícia Valéria Castelo-Branco, Israel Higino de Sousa, Ana Paula Azevedo-Santos, André Alvares Marques Vale, Silvio Gomes Monteiro, Rossy-Eric Pereira Soares, Sulayne Janayna Araujo Guimarães, Jessyane Rodrigues do Nascimento, Silma Regina Ferreira Pereira
Considering the occurrence of interactions between flavonoids and FBS proteins, mainly albumin (Liu et al.2010, Hu et al. 2012, Pal and Saha 2014, Greenwell and Rahman 2015, Tang et al.2017, Genget al.2018, Fujiiet al.2019), we initially investigated the effect of FBS on the DCMF cytotoxic activity. We carried out the sulforhodamine B colorimetric assay (SRB) as described by Vichai and Kirtikara (2006). Firstly, the DCMF solution was prepared by dissolving 3 mg of DCMF in 100 μL of dimethylsulfoxide -DMSO (Sigma-Aldrich; USA). Preliminary tests were conducted to ascertain the protocol used in the cytotoxicity assays. Initially, the cells were exposed to concentrations ranging from 1.5 to 100 μg/mL of DCMF for 24 and 48 h. As a result, more than 90% of the cells became nonviable when exposed to >20 μg/mL. Consequently, treatments with 1, 5, 10, and 20 μg/mL for 48 h were established. Untreated cells and cells exposed to the highest concentration of DMSO in the culture medium (0.07%) were used as negative and vehicle control, respectively.