scholarly journals TMEM16F Regulates Baseline Phosphatidylserine Exposure and Cell Viability in Human Embryonic Kidney Cells

2016 ◽  
Vol 38 (6) ◽  
pp. 2452-2463 ◽  
Author(s):  
Laura K. Schenk ◽  
Ulf Schulze ◽  
Sebastian Henke ◽  
Thomas Weide ◽  
Hermann Pavenstädt
Keyword(s):  
Signaling Pathways ◽  
Transmembrane Protein ◽  
Cell Cycle Control ◽  
Akt Signaling ◽  
Hek293 Cells ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Human Embryonic Kidney Cells ◽  
Phospholipid Scramblase ◽  
Phosphatidylserine Exposure

Background / Aims: TMEM16F is a transmembrane protein from a conserved family of Ca2+-activated proteins, which is highly expressed in several tissues. TMEM16F confers phospholipid scramblase activity and Ca2+-activated electrolyte channel activity. Potentially thereby, TMEM16F is involved in cell cycle control and apoptotic signaling. The present study evaluated the role of TMEM16F on cell proliferation and viability in Human Embryonic Kidney cells. Methods: An inducible knockdown of TMEM16F was generated and markers of apoptosis and proliferation were assessed via flow cytometry, western blotting and MTT uptake assay under different conditions. Results: TMEM16F knockdown resulted in attenuated growth of HEK293 cells. This observation correlated with an increased phosphatidylserine exposure and a decreased fraction of viable cells. Interestingly, the cells were not sensitized to Staurosporine- induced cell death. Western blot analyses displayed a parallel activation of pro- and antiapoptotic signaling pathways: Caspase 3 cleavage and Cyclin D1 abundance were simultaneously increased. Furthermore, knockdown of TMEM16F led to activation of AKT signaling. Conclusion: TMEM16F modifies viability of Human Embryonic Kidney cells via its function as a phospholipid scramblase and activation of AKT signaling pathways.

scholarly journals Heme oxygenase-2 (HO-2) binds and buffers labile heme, which is largely oxidized, in human embryonic kidney cells

2021 ◽  
Author(s):  
David A Hanna ◽  
Courtney M Moore ◽  
Liu Liu ◽  
Xiaojing Yuan ◽  
Angela S Fleischhacker ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Physiological Function ◽  
Hek293 Cells ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Human Embryonic Kidney Cells ◽  
Heme Oxygenases ◽  
Heme Trafficking ◽  
Ferrous Heme

Heme oxygenases (HO) detoxify heme by oxidatively degrading it into carbon monoxide, iron, and biliverdin, which is reduced to bilirubin and excreted. Humans express two isoforms: inducible HO-1, which is up-regulated in response to various stressors, including excess heme, and constitutive HO-2. While much is known about the regulation and physiological function of HO-1, comparatively little is known about the role of HO-2 in regulating heme homeostasis. The biochemical necessity for expressing constitutive HO-2 is largely dependent on whether heme is sufficiently abundant and accessible as a substrate under conditions in which HO-1 is not induced. By measuring labile heme, total heme, and bilirubin in human embryonic kidney HEK293 cells with silenced or over-expressed HO-2, and various HO-2 mutant alleles, we found that endogenous heme is too limiting to support HO-2 catalyzed heme degradation. Rather, we discovered that a novel role for HO-2 is to bind and buffer labile heme. Taken together, in the absence of excess heme, we propose that HO-2 regulates heme homeostasis by acting as a heme buffering factor in control of heme bioavailability. When heme is in excess, HO-1 is induced and both HO-2 and HO-1 can provide protection from heme toxicity by enzymatically degrading it. Our results explain why catalytically inactive mutants of HO-2 are cytoprotective against oxidative stress. Moreover, the change in bioavailable heme due to HO-2 overexpression, which selectively binds ferric over ferrous heme, is consistent with the labile heme pool being oxidized, thereby providing new insights into heme trafficking and signaling.

scholarly journals Transcriptomic and Proteomic Analysis Reveals Mechanisms of Patulin-Induced Cell Toxicity in Human Embryonic Kidney Cells

Toxins ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 681
Author(s):  
Nianfa Han ◽  
Ruilin Luo ◽  
Jiayu Liu ◽  
Tianmin Guo ◽  
Jiayu Feng ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Digital Gene Expression ◽  
Hek293 Cells ◽  
Kidney Cells ◽  
Oxygen Species ◽  
Human Embryonic Kidney ◽  
Absolute Quantitation ◽  
Human Embryonic Kidney Cells ◽  
Isobaric Tags ◽  
Induced Apoptosis

Patulin (PAT) is a natural mycotoxin that commonly contaminates fruits and fruit-based products. Previous work indicated that PAT-induced apoptosis in which reactive oxygen species (ROS) are involved in human embryonic kidney (HEK293) cells. To uncover novel aspects of the possible mechanism of PAT nephrotoxicity, the transcriptome and proteome profiles were investigated using the digital gene expression (DGE) and isobaric tags for relative and absolute quantitation (iTRAQ) proteomic approaches. A total of 127 genes and 85 proteins were found to express differentially in response to 5 μM PAT for 10 h in HEK293 cells. The most dramatic changes of expression were noticed with genes or proteins related to apoptosis, oxidative phosphorylation ribosome and cell cycle. Especially, the activation of caspase 3, UQCR11, active transport form and endocytosis appeared to be crucial in PAT kidney cytotoxicity. PAT also seemed to be associated with cancer and neuropathic disease as pathways associated with carcinogenesis, Alzheimer’s disease and Parkinson’s disease were induced. Overall, this study served to uncover overall insights associated with signaling pathway that modulated the PAT toxicity mechanism.

scholarly journals Functional Comparison between VP64-dCas9-VP64 and dCas9-VP192 CRISPR Activators in Human Embryonic Kidney Cells

2021 ◽  
Vol 22 (1) ◽  
pp. 397
Author(s):  
Nasir Javaid ◽  
Thuong L. H. Pham ◽  
Sangdun Choi
Keyword(s):  
Protein Level ◽  
Reference Genes ◽  
Mrna Level ◽  
High Specificity ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Human Embryonic Kidney Cells ◽  
Sox2 Expression

Reversal in the transcriptional status of desired genes has been exploited for multiple research, therapeutic, and biotechnological purposes. CRISPR/dCas9-based activators can activate transcriptionally silenced genes after being guided by gene-specific gRNA(s). Here, we performed a functional comparison between two such activators, VP64-dCas9-VP64 and dCas9-VP192, in human embryonic kidney cells by the concomitant targeting of POU5F1 and SOX2. We found 22- and 6-fold upregulations in the mRNA level of POU5F1 by dCas9-VP192 and VP64-dCas9-VP64, respectively. Likewise, SOX2 was up-regulated 4- and 2-fold using dCas9-VP192 and VP64dCas9VP64, respectively. For the POU5F1 protein level, we observed 3.7- and 2.2-fold increases with dCas9-VP192 and VP64-dCas9-VP64, respectively. Similarly, the SOX2 expression was 2.4- and 2-fold higher with dCas9-VP192 and VP64-dCas9-VP64, respectively. We also confirmed that activation only happened upon co-transfecting an activator plasmid with multiplex gRNA plasmid with a high specificity to the reference genes. Our data revealed that dCas9-VP192 is more efficient than VP64-dCas9-VP64 for activating reference genes.

Fenton Reaction-Generated Advanced Oxidation Protein Products Induces Inflammation in Human Embryonic Kidney Cells

Inflammation ◽  
2016 ◽  
Vol 39 (4) ◽  
pp. 1285-1290 ◽  
Author(s):  
Guilherme Vargas Bochi ◽  
Vanessa Dorneles Torbitz ◽  
Roberto Christ Vianna Santos ◽  
Monica Cubillos-Rojas ◽  
José Luis Rosa López ◽  
...  
Keyword(s):  
Fenton Reaction ◽  
Advanced Oxidation ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Advanced Oxidation Protein Products ◽  
Human Embryonic Kidney Cells

scholarly journals Human embryonic kidney cells: stable transformation with an origin-defective simian virus 40 DNA and use as hosts for human papovavirus replication.

1984 ◽  
Vol 4 (2) ◽  
pp. 379-382 ◽  
Author(s):  
E O Major ◽  
P Matsumura
Keyword(s):  
Dna Replication ◽  
Sequence Data ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Human Embryonic Kidney Cells ◽  
Defective Mutant ◽  
Dna Replication Origin ◽  
Human Papovavirus

An origin-defective mutant DNA of simian virus 40 immortalized human embryonic kidney cells, maintaining a T protein which could function for human papovavirus BK DNA replication but not for human papovavirus JC DNA replication. Neither BK virions nor capsid proteins were produced in these cells. This may indicate that the simian virus 40 T protein in human embryonic kidney cells is competent for maintaining transformation and initiating and completing DNA replication for BK but is not competent for switching to late gene functions. Furthermore, it appears that the JC DNA replication origin cannot efficiently use the simian virus 40 T protein for its DNA synthesis, as suggested by its DNA sequence data (R. Frisque, J. Virol. 46:170-176, 1983; T. Miyamura, H. Jikoya, E. Soeda, and K. Yoshiike, J. Virol. 45:73-79, 1983).

A smart rhodamine–pyridine conjugate for bioimaging of thiocyanate in living cells

RSC Advances ◽  
2015 ◽  
Vol 5 (125) ◽  
pp. 103350-103357 ◽  
Author(s):  
Sandip Mandal ◽  
Animesh Sahana ◽  
Arnab Banerjee ◽  
Damir A. Safin ◽  
Maria G. Babashkina ◽  
...  
Keyword(s):  
Visible Light ◽  
Living Cells ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Eye Detection ◽  
Human Embryonic Kidney Cells ◽  
Naked Eye ◽  
Naked Eye Detection

A rhodamine–pyridine conjugate, REDA-2PC, can selectively monitor NCS− in human embryonic kidney cells 293. Visible light excitable probe allows fluorescence and naked eye detection of nanomolar NCS−.

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