ColonyCountJ: A User-Friendly Image J Add-on Program for Quantification of Different Colony Parameters in Clonogenic Assay

Glioblastoma multiform (GBM) is a type of aggressive brain cancer with limited success in standard treatment. MicroRNAs are one of the most beneficial tools for diagnosis, prognosis, and treatment of cancer. This study aimed to investigate the effect of miR‐579 on cellular behaviors and expression of PI3K/AKT signaling pathway in GBM cell lines. In the present study, miR‐579 was overexpressed in U251 and A‐172 cell lines by using lentil vector, and its effect on cellular behavior such as proliferation and migration was investigated by the cell cycle, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT), Annexin V, colony formation, Transwell and wound healing assays. MiR‐579 predicted target genes (AKT1, Rheb, PDK1, and a few others) were also evaluated by real‐time polymerase chain reaction or luciferase assay and Western blot analysis. Our results represented that overexpression of miR‐579 could inhibit proliferation, migration, cell cycle and also promoted the apoptosis of GBM cell lines. The luciferase reporter assay showed miR‐579 directly targets the 3 UTR of mTOR, Rheb, and PDK1 and repressed their expressions. Furthermore, the Western blot analysis showed that miR‐579 could downregulate the AKT1 and Rheb protein expression. Overall, our findings propose that miR‐579 functions as a novel tumor suppressor gene in GBM by regulating the PI3K/AKT signaling pathway and may serve as a therapeutic target for clinical therapy of glioblastoma multiform.

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“…ColonyCount J, an Image J Add-on Program, determined the colony formation rate. 17 All experiments were performed in triplicate.…”

Section: Colony Formation Assaymentioning

confidence: 99%

The effect of miR‐579 on the PI3K/AKT pathway in human glioblastoma PTEN mutant cell lines

Kalhori

Irani

Soleimani

et al. 2019

J of Cellular Biochemistry

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“…Developed colonies were fixed with methanol and acetic acid and stained using 0.4% trypan blue (207-03252; Wako). Colonies were quantified by ColonyCountJ 25 .…”

Section: Methodsmentioning

confidence: 99%

Calcineurin-mediated dephosphorylation enhances the stability and transactivation of c-Myc

Masaki,

Habara,

Hanaki

et al. 2023

Sci Rep

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Abstractc-Myc, a transcription factor, induces cell proliferation and is often aberrantly or highly expressed in cancers. However, molecular mechanisms underlying this aberrantly high expression remain unclear. Here, we found that intracellular Ca2+ concentration regulates c-Myc oncoprotein stability. We identified that calcineurin, a Ca2+-dependent protein phosphatase, is a positive regulator of c-Myc expression. Calcineurin depletion suppresses c-Myc targeted gene expression and c-Myc degradation. Calcineurin directly dephosphorylates Thr58 and Ser62 in c-Myc, which inhibit binding to the ubiquitin ligase Fbxw7. Mutations within the autoinhibitory domain of calcineurin, most frequently observed in cancer, may increase phosphatase activity, increasing c-Myc transcriptional activity in turn. Notably, calcineurin inhibition with FK506 decreased c-Myc expression with enhanced Thr58 and Ser62 phosphorylation in a mouse xenograft model. Thus, calcineurin can stabilize c-Myc, promoting tumor progression. Therefore, we propose that Ca2+ signaling dysfunction affects cancer-cell proliferation via increased c-Myc stability and that calcineurin inhibition could be a new therapeutic target of c-Myc-overexpressing cancers.

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“…However, the clonogenic cell survival assay is tedious and time-consuming and offers very low sample throughput. [165,166] As such, metabolic-based assays (e.g. MTT or CellTiter-Glo®) can in part overcome these limitations and are often used as a first screening approach to identify reasonable NP concentration and ionizing radiation dose ranges to probe novel combined treatment modalities on the in vitro level, followed by the more exact clonogenic cell survival assay.…”

Section: Reactivation Of An Antitumor Immune Responsementioning

confidence: 99%

Prospects of Nanoparticle-based Radioenhancement for Radiotherapy

Gerken

Gerdes

Pruschy

et al. 2023

Preprint

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Radiotherapy is a key pillar of solid cancer treatment. Despite high level of conformal dose deposition, radiotherapy is limited due to co-irradiation of organs-at risk and subsequent normal tissue toxicities. Nanotechnology offers an attractive opportunity for increasing the efficacy and safety of cancer radiotherapy. Leveraging the freedom of design and the growing synthetic capabilities of the nanomaterial-community, a variety of engineered nanomaterials have been designed and investigated as radiosensitizers or radioenhancers. While research so far has been primarily focused on gold nanoparticles and other high atomic number materials to increase the absorption cross section of tumor tissue, recent studies are challenging the traditional concept of high-Z nanoparticle radioenhancers and highlight the importance of catalytic activity. This review provides a concise overview on the fundamental knowledge of nanoparticle radioenhancement mechanisms and their quantification. It critically discusses potential radioenhancer candidate materials and general design criteria for different radiation therapy modalities, and concludes with research priorities in order to advance the development of nanomaterials, to enhance the efficacy of radiotherapy and to increase at the same time the therapeutic window.

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ColonyCountJ: A User-Friendly Image J Add-on Program for Quantification of Different Colony Parameters in Clonogenic Assay

Cited by 13 publications

References 6 publications

The effect of miR‐579 on the PI3K/AKT pathway in human glioblastoma PTEN mutant cell lines

The effect of miR‐579 on the PI3K/AKT pathway in human glioblastoma PTEN mutant cell lines

Calcineurin-mediated dephosphorylation enhances the stability and transactivation of c-Myc

Prospects of Nanoparticle-based Radioenhancement for Radiotherapy

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