DRAQ7 as an Alternative to MTT Assay for Measuring Viability of Glioma Cells Treated With Polyphenols

Rooprai, Harcharan K.; Lawrence, Patrick Varun; Keshavarz, Sholeh; Yashod, Pooja; Gullan, Richard; Selway, Richard; Davies, Derek

doi:10.21873/anticanres.14553

Cited by 11 publications

(8 citation statements)

References 33 publications

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“…To quantify cell death, the cells were stained with DRAQ7, a membrane-impermeable dye that stains the double-stranded DNA of dead cells. 42 The cells were cultured for 3 days in the presence or absence of the degradation products digested with PK. By studying cell toxicity, we found that the degradation products of either uncrosslinked or crosslinked scaffolds did not affect the cell number or cell death in human dermal fibroblasts (Fig.…”

Section: Resultsmentioning

confidence: 99%

Degradation products of crosslinked silk fibroin scaffolds modulate the immune response but not cell toxicity

et al. 2023

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Section: Resultsmentioning

confidence: 99%

Degradation products of crosslinked silk fibroin scaffolds modulate the immune response but not cell toxicity

et al. 2023

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“…Glioma is a primary intracranial tumor with high incidence and high degree of malignancy, and the average survival time of patients is relatively short [12]. Rab23 gene is closely related to the progression of cancer.…”

Section: Discussionmentioning

confidence: 99%

Effect of Licochalcone-A Combined with Rab23 Gene on Proliferation of Glioma U251 Cells

Dong

Cui

et al. 2022

Evidence-Based Complementary and Alternative Medicine

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This research aimed to explore the effect of Licochalcone-A (LCA) combined with Rab23 gene on the proliferation, migration, and invasion of glioma U251 cells through the Wnt/β-catenin signaling pathway. The glioma U251 cell line was taken as the research object, and the Rab23 overexpression plasmid was constructed. According to the treatment method, U251 cells were rolled into blank control group (BC), Rab23 overexpression plasmid transfection group (Rab23), 25 μmol·L−1 LCA treatment group (LCA), and Rab23 overexpression plasmid transfection combined with 25 μmol·L−1 LCA treatment group (Rab23 + LCA). Subsequently, the ability of cell proliferation, migration, and invasion of each group was detected by methyl thiazolyl tetrazolium (MTT) assay, scratch healing test, and Transwell cell invasion test, respectively. Western blot was implemented to detect the expression differences of cell proliferation antigen Ki-67, apoptosis-related proteins Bcl-2 and Bax, and Wnt/β-catenin pathway-related proteins β-catenin, glycogen synthase kinase-3 (GSK3β), Axin2, and c-myc. The results showed the successful construction of Rab23 overexpression and stable transfection U251 cell line. After grouping and treatments, the cell proliferation, migration, and invasion ability of the Rab23 group, LCA group, and Rab23 + LCA group was substantially reduced relative to BC group ( P < 0.05 ). In addition, the cell proliferation, migration, and invasion ability of Rab23 + LCA group decreased relatively more significantly. The expression levels of Ki-67, Bcl-2, β-catenin, and c-myc in the Rab23, LCA, and Rab23 + LCA groups were greatly lower versus those of BC group. Moreover, the protein expression levels of Bax, GSK3β, and Axin2 were considerably increased ( P < 0.05 ), while the expression of protein in Rab23 + LCA group increased notably. These findings indicate that LCA combined with Rab23 gene can inhibit the proliferation, migration, and invasion of glioma U251 cells through the Wnt/β-catenin signaling and can promote cell apoptosis.

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“…The co-delivery complexes have been well characterized in terms of pDNA CC, shape, mean size, surface charges and biocompatibility At this stage, we must also mention that the cellular viability data, presented in Figure 4, does not fully correlate with both the quantified p53 levels (Table 2) and the observed caspase-3 activity in HeLa cells. For this discrepancy can, probably, contribute the most common reported limitations of MTT assay, such as, lack of sensitivity, interference with cell culture medium and overestimation of cell viability [63,64].…”

Section: Discussionmentioning

confidence: 99%

Development of Tailor-Made Dendrimer Ternary Complexes for Drug/Gene Co-Delivery in Cancer

et al. 2021

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Cancer gene therapy, mediated by non-viral systems, remains a major research focus. To contribute to this field, in this work we reported on the development of dendrimer drug/gene ternary complexes. This innovative approach explored the great capacity of both polyamidoamine (PAMAM)-paclitaxel (PTX) conjugate and polyethylenimine (PEI) polymers to complex a p53-encoding plasmid DNA (pDNA), highlighting the utility of considering two compacting agents. The pDNA complexation capacity has been investigated as function of the nitrogen to phosphate groups ratio (N/P), which revealed to be a tailoring parameter. The physicochemical properties of the conceived ternary complexes were revealed and were found to be promising for cellular transfection. Furthermore, the formulated co-delivery systems demonstrated to be biocompatible. The ternary systems were able of cellular internalization and payload intracellular release. Confocal microscopy studies showed the co-localization of stained pDNA with the nucleus of cancer cells, after transfection mediated by these carriers. From this achievement, p53 gene expression occurred with the production of protein. Moreover, the activation of caspase-3 indicated apoptosis of cancer cells. This work represents a great progress on the design of dendrimer drug/gene co-delivery systems towards a more efficient cancer therapy. In this way, it instigates further in vitro studies concerning the evaluation of their therapeutic potential, expectedly supported by the synergistic effect, in tumoral cells.

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DRAQ7 as an Alternative to MTT Assay for Measuring Viability of Glioma Cells Treated With Polyphenols

Cited by 11 publications

References 33 publications

Degradation products of crosslinked silk fibroin scaffolds modulate the immune response but not cell toxicity

Degradation products of crosslinked silk fibroin scaffolds modulate the immune response but not cell toxicity

Effect of Licochalcone-A Combined with Rab23 Gene on Proliferation of Glioma U251 Cells

Development of Tailor-Made Dendrimer Ternary Complexes for Drug/Gene Co-Delivery in Cancer

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