scholarly journals Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

2015 ◽  
Vol 112 (27) ◽  
pp. 8344-8349 ◽  
Author(s):  
Charles W. Morgan ◽  
Juan E. Diaz ◽  
Samantha G. Zeitlin ◽  
Daniel C. Gray ◽  
James A. Wells
Keyword(s):  
Protein Function ◽  
Human Cancer ◽  
Gene Replacement ◽  
Transcriptional Level ◽  
Human Cancer Cell Lines ◽  
Cooperative Action ◽  
Dna Laddering ◽  
Caspase Activated Dnase ◽  
Executioner Caspases

Cellular demolition during apoptosis is completed by executioner caspases, that selectively cleave more than 1,500 proteins but whose individual roles are challenging to assess. Here, we used an optimized site-specific and inducible protease to examine the role of a classic apoptotic node, the caspase-activated DNase (CAD). CAD is activated when caspases cleave its endogenous inhibitor ICAD, resulting in the characteristic DNA laddering of apoptosis. We describe a posttranscriptional gene replacement (PTGR) approach where endogenous biallelic ICAD is knocked down and simultaneously replaced with an engineered allele that is susceptible to inducible cleavage by tobacco etch virus protease. Remarkably, selective activation of CAD alone does not induce cell death, although hallmarks of DNA damage are detected in human cancer cell lines. Our data strongly support that the highly cooperative action of CAD and inhibition of DNA repair systems are critical for the DNA laddering phenotype in apoptosis. Furthermore, the PTGR approach provides a general means for replacing wild-type protein function with a precisely engineered mutant at the transcriptional level that should be useful for cell engineering studies.

scholarly journals CRISPR enriches for cells with mutations in a p53-related interactome, and this can be inhibited

2021 ◽  
Author(s):  
Long Jiang ◽  
Katrine Ingelshed ◽  
Yunbing Shen ◽  
Sanjaykumar V. Boddul ◽  
Vaishnavi Srinivasan Iyer ◽  
...  
Keyword(s):  
Cellular Response ◽  
Human Cancer ◽  
Genetic Alterations ◽  
Dna Breaks ◽  
Data Set ◽  
Human Cancer Cell Lines ◽  
Cycle Arrest ◽  
Double Stranded Dna ◽  
A Cell

CRISPR/Cas9 can be used to inactivate or modify genes by inducing double-stranded DNA breaks1–3. As a protective cellular response, DNA breaks result in p53-mediated cell cycle arrest and activation of cell death programs4,5. Inactivating p53 mutations are the most commonly found genetic alterations in cancer, highlighting the important role of the gene6–8. Here, we show that cells deficient in p53, as well as in genes of a core CRISPR-p53 tumor suppressor interactome, are enriched in a cell population when CRISPR is applied. Such enrichment could pose a challenge for clinical CRISPR use. Importantly, we identify that transient p53 inhibition suppresses the enrichment of cells with these mutations. Furthermore, in a data set of >800 human cancer cell lines, we identify parameters influencing the enrichment of p53 mutated cells, including strong baseline CDKN1A expression as a predictor for an active CRISPR-p53 axis. Taken together, our data identify strategies enabling safe CRISPR use.

scholarly journals Role of ATF5 in the invasive potential of diverse human cancer cell lines

2016 ◽  
Vol 474 (3) ◽  
pp. 509-514 ◽  
Author(s):  
Akihiro Nukuda ◽  
Hiroki Endoh ◽  
Motoaki Yasuda ◽  
Takeomi Mizutani ◽  
Kazushige Kawabata ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
Human Cancer ◽  
Cancer Cell Lines ◽  
Human Cancer Cell ◽  
Invasive Potential ◽  
Human Cancer Cell Lines

scholarly journals Cytotoxicity evaluation of a new series of naphthalimide derivatives against human cancer cell lines

2019 ◽  
Author(s):  
Alireza Aliabadi ◽  
Ahmad Mohammadi-Farani ◽  
Arash Haqiqi ◽  
Elham Khanlari
Keyword(s):  
Cell Lines ◽  
Human Cancer ◽  
Basic Study ◽  
Human Cancer Cell Lines ◽  
Lead Compounds ◽  
Cancerous Cell ◽  
Pc3 Cells ◽  
New Anticancer Drugs ◽  
Ht 29

Introduction: Cancer has been known as one the main causes of the death in the world. In recent researches, discovery of effective, selective and safe medications is a priority and emergency. In recent reports, the role of lipoxygenases (LOX) has been confirmed in neoplastic diseases. Some isoenzymes such as 5, 12 and 15 have more importance in the neoplasia. According to the efficacy of naphthalimides as LOX inhibitor, herein we explored the cytotoxicity of naphthalimide derivatives.  Methods: A basic study was carried out in the current research. The cytotoxicity of a new series of naphthalimide-based 15-LOX-1 inhibitors was evaluated in three cancerous cell lines namely SKNMC (neuroblastoma), PC3 (prostatic cancer), HT29 (colorectal cancer) using MTT protocol and the obtained data was compared to doxorubicin. Calculation of the IC50 of tested compounds was performed by regression analysis using Prism-6 software. Results: Totally, all tested compounds exhibited inferior activity than doxorubicin towards HT-29, SKNMC and PC3 cell lines. SKNMC cell line rendered more sensitivity to tested compounds. Amongst the compounds 3a-3m, compound 3e (3-methoxy) and 3i (4-F) with IC50 = 5.92±1.78 µM and 10.04±1.7 µM were the most potent derivatives towards PC3 cells. Conclusion: Although all compounds did not exert more potency in comparison with doxorubicin, some of them showed remarkable cytotoxicity. The potent derivatives could be introduced as novel lead compounds for development of new anticancer drugs.

881Modulation of osteoinductive properties of human cancer cell lines: Essential role of noggin

2005 ◽  
Vol 4 (3) ◽  
pp. 223
Author(s):  
G. Thalmann ◽  
R. Schwaninger ◽  
C. Rentsch ◽  
A. Wetterwald ◽  
F. Geissbühler ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
Essential Role ◽  
Human Cancer ◽  
Cancer Cell Lines ◽  
Human Cancer Cell ◽  
Human Cancer Cell Lines

scholarly journals Conclusive Evidence for OCT4 Transcription in Human Cancer Cell Lines: Possible Role of a Small OCT4-Positive Cancer Cell Population

Stem Cells ◽  
2018 ◽  
Vol 36 (9) ◽  
pp. 1341-1354 ◽  
Author(s):  
Tomoyuki Miyamoto ◽  
Nobuhiko Mizuno ◽  
Mitsuko Kosaka ◽  
Yoko Fujitani ◽  
Eiji Ohno ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
Cell Population ◽  
Human Cancer ◽  
Cancer Cell Lines ◽  
Conclusive Evidence ◽  
Human Cancer Cell ◽  
Human Cancer Cell Lines ◽  
Cancer Cell Population

scholarly journals Systematic expression and bioinformatics analysis of fibrinogen-like protein 1 in human cancer and its co-expression network

2021 ◽  
Author(s):  
Qian Wang ◽  
Shu-zhen Chen ◽  
Hua-wei Zhang
Keyword(s):  
Hepatocellular Carcinoma ◽  
Immune Response ◽  
Bioinformatics Analysis ◽  
Human Cancer ◽  
Transcriptional Level ◽  
Human Cell Lines ◽  
Human Carcinoma ◽  
Pathway Enrichment ◽  
Coexpression Networks

Abstract Background Fibrinogen-like protein 1 was initially found in a study on hepatocellular carcinoma and it is overexpressed in human cell lines and rats. Recently, investigations have focused on fibrinogen-like protein 1 dysfunction in carcinogenesis. Our study aims to determine the role of fibrinogen-like protein 1 and its possible role in human carcinoma. Methods Fibrinogen-like protein 1 expression in different neoplasms was assessed by Oncomine. Fibrinogen-like protein 1 coexpression networks in various cancers were established using Coexpedia. Finally, we investigated the potential functions of fibrinogen-like protein 1 with gene ontology and pathway enrichment analyses with the FunRich V3. Results Fibrinogen-like protein 1 was overexpressed in several kinds of neoplasms at the transcriptional level. Coexpression networks showed that fibrinogen-like protein 1 regulates immune response and lipid related pathways. Conclusions The present results offer the possibility that fibrinogen-like protein 1 acts as a therapeutic target for some types of cancers and may take part in carcinogenesis.

scholarly journals Regulation of let-7 microRNAs by a two-step turnover pathway is of critical pathophysiological significance in humans

2021 ◽  
Author(s):  
Rohit Nalawade ◽  
Tamoghna Chowdhury ◽  
Saibal Chatterjee
Keyword(s):  
Cellular Proliferation ◽  
Human Cancer ◽  
Metastatic Potential ◽  
Mirna Biogenesis ◽  
Human Cancer Cell Lines ◽  
Physiological Processes ◽  
Consistent Manner ◽  
Transcriptomics Data ◽  
And Function

MicroRNAs (miRNAs) are critical regulators of diverse developmental and physiological processes in animals, and their dysregulation has been linked to various disorders and diseases. Not only multiple regulatory mechanisms acting at different levels of human miRNA biogenesis determine miRNA abundance and function, but a few recently identified factors by facilitating miRNA turnover also make critical contributions. We demonstrate that the ribonuclease XRN2, whose worm ortholog had previously been shown to actively degrade let-7 family of miRNAs, can degrade the mature forms of certain let-7 family members in multiple human cancer cell lines, without affecting their precursors. The XRN2-mediated turnover of let-7 has patho-physiological significance as XRN2 depletion results in a reduction in the expression of a number of oncogenes, and diminishes the proliferative and metastatic potential of cancer cells. The clinical relevance of these observations is also verified in tumour transcriptomics data from public RNA-sequencing datasets, where we observe that higher XRN2 mRNA expression is inversely correlated with the levels of mature let-7 miRNAs and associated with poor survival in hepatocellular carcinoma, lung adenocarcinoma, and glioblastoma. We also demonstrate that miRNA turnover is a step-wise process, where a miRNA is released from the grasp of Argonaute before its degradation. This yet unidentified miRNA releasing factor is proteinaceous in nature and its activity is kinetically linked with XRN2-mediated turnover of miRNAs. Our analyses of the patient-derived transcriptomics data also show that XRN2, via its regulation of let-7, affects pathways related to cellular proliferation, development, and signalling in a consistent manner across epithelial and glial cell lineages. Collectively, our studies suggest an important role of XRN2 in regulating cancer physiology through degradation of the let-7 family of miRNAs.

scholarly journals Bioactive Brominated Oxindole Alkaloids from the Red Sea Sponge Callyspongia siphonella

Marine Drugs ◽  
2019 ◽  
Vol 17 (8) ◽  
pp. 465 ◽  
Author(s):  
Seham S. El-Hawary ◽  
Ahmed M. Sayed ◽  
Rabab Mohammed ◽  
Hossam M. Hassan ◽  
Mostafa E. Rateb ◽  
...  
Keyword(s):  
Cytotoxic Effect ◽  
Inhibitory Concentration ◽  
Human Cancer ◽  
Human Cancer Cell ◽  
Human Cancer Cell Lines ◽  
Gram Positive Bacteria ◽  
Fouling Organisms ◽  
Oxindole Alkaloids

In the present study, LC-HRESIMS-assisted dereplication along with bioactivity-guided isolation led to targeting two brominated oxindole alkaloids (compounds 1 and 2) which probably play a key role in the previously reported antibacterial, antibiofilm, and cytotoxicity of Callyspongia siphonella crude extracts. Both metabolites showed potent antibacterial activity against Gram-positive bacteria, Staphylococcus aureus (minimum inhibitory concentration (MIC) = 8 and 4 µg/mL) and Bacillus subtilis (MIC = 16 and 4 µg/mL), respectively. Furthermore, they displayed moderate biofilm inhibitory activity in Pseudomonas aeruginosa (49.32% and 41.76% inhibition, respectively), and moderate in vitro antitrypanosomal activity (13.47 and 10.27 µM, respectively). In addition, they revealed a strong cytotoxic effect toward different human cancer cell lines, supposedly through induction of necrosis. This study sheds light on the possible role of these metabolites (compounds 1 and 2) in keeping fouling organisms away from the sponge outer surface, and the possible applications of these defensive molecules in the development of new anti-infective agents.

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