Melanopsin Signaling Pathway in HEK293 Cell Line with Stable Expression of Human Melanopsin: Possible Participation of Phospholipase C beta 4 and Diacylglycerol

The adenoviral E1B-55k protein present in HEK293 cells mediates abnormal accumulation of key WNT signaling proteins in large cytoplasmic aggregates

10.21203/rs.3.rs-840397/v1 ◽

2021 ◽

Author(s):

Petter Angell Olsen ◽

Stefan Krauss

Keyword(s):

Wnt Signaling ◽

Cell Line ◽

Signaling Pathway ◽

Transcriptional Activation ◽

Cellular Localization ◽

Hek293 Cell ◽

Wnt Signaling Pathway ◽

Hek293 Cells ◽

Protein Levels ◽

Hek293 Cell Line

Abstract HEK293 cells are one of the most widely used cell lines in research and HEK293 cells are frequently used as an in vitro model for studying the WNT signaling pathway. The HEK293 cell line was originally established by transfection of human embryonic kidney cells with sheared adenovirus 5 DNA and it is known that that HEK293 cells stably express the adenoviral E1A and E1B-55k proteins. Here we show that HEK293 cells display an unexpected distribution of key components of the WNT/β-catenin signaling pathway where AXIN1, APC, DVL2 and tankyrase are all co-localized in large spherical cytoplasmic aggregates. The cytoplasmic aggregates are enclosed by a narrow layer of the adenoviral E1B-55k protein. Reduction of E1B-55k protein levels leads to disappearance of the cytoplasmic aggregates thus corroborating an essential role of the E1B-55k protein in mediating the formation of the aggregates. Furthermore, HEK293 cells with reduced E1B-55k protein levels display reduced levels of transcriptional activation of WNT/β-catenin signaling upon stimulation by the Wnt3A agonist. The demonstrated influence of the E1B-55k protein on the cellular localization of WNT/β-catenin signaling components and on transcriptional regulation of WNT/β-catenin signaling asks for caution in the interpretation of data derived from the HEK293 cell line.

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Lemon Juice Mediated Synthesis of 3-Substituted Quinazolin-4(3H)-Ones and their Pharmacological Evaluation

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520619666190723151909 ◽

2020 ◽

Vol 19 (16) ◽

pp. 2001-2009 ◽

Cited By ~ 1

Author(s):

Malavattu G. Prasad ◽

C. Vijaya Lakshmi ◽

Naresh K. Katari ◽

Sreekantha B. Jonnalagadda ◽

Manojit Pal

Keyword(s):

Cell Line ◽

Cell Lines ◽

Hek293 Cell ◽

Glyoxylic Acid ◽

Lemon Juice ◽

Cytotoxic Agents ◽

Diverse Range ◽

Hek293 Cell Line ◽

Three Component Reaction ◽

Mcf 7

Background: Compounds containing the quinazoline-4(3H)-one framework constitute an important class of fused N-heterocycles that are found in more than 200 naturally occurring alkaloids. These compounds also show a diverse range of pharmacological activities including antitumor properties. This prompted us to explore a series of quinazolin-4-(3H)-one derivatives having no substituent at C-2 as potential cytotoxic agents. Objective: The objective of this study was to synthesize and evaluate 3-substituted quinazolin-4(3H)-one derivatives for their potential cytotoxic properties. Methods: A convenient method has been developed for the rapid synthesis of this class of compounds under a mild and non-hazardous reaction condition in good yields. The methodology involved a three-component reaction employing isatoic anhydride, amines and glyoxylic acid as reactants in the presence of lemon juice in PEG- 400 at room temperature (25-30ºC) under ultrasound irradiation. All the synthesized compounds were screened via an MTT assay for their potential cytotoxic properties in vitro using the cancerous cell lines e.g. A549, A2780, HepG2, K562, MCF-7 and HCT-116 and a non-cancerous HEK293 cell line. Results: Several compounds such as 3a, 3b, 3d, 3e and 3f showed promising growth inhibition against these cancer cell lines but no significant effects on HEK293 cell line. The IC50 values of these compounds were comparable to doxorubicin whereas 3f significantly induced apoptosis in MCF-7 cells that also was comparable to doxorubicin. Conclusion: An ultrasound-assisted MCR facilitated by lemon juice has been developed to synthesize 3- substituted quinazolin-4(3H)-one derivatives that could act as potential anticancer agents.

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Affecting HEK293 Cell Growth and Production Performance by Modifying the Expression of Specific Genes

Cells ◽

10.3390/cells10071667 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1667

Author(s):

Laura Abaandou ◽

David Quan ◽

Joseph Shiloach

Keyword(s):

Cell Line ◽

Cell Lines ◽

Hek293 Cell ◽

Chinese Hamster ◽

Production Performance ◽

Cellular Processes ◽

Hek293 Cell Line ◽

Global Engineering ◽

Genomic Tools ◽

Serum Free Suspension

The HEK293 cell line has earned its place as a producer of biotherapeutics. In addition to its ease of growth in serum-free suspension culture and its amenability to transfection, this cell line’s most important attribute is its human origin, which makes it suitable to produce biologics intended for human use. At the present time, the growth and production properties of the HEK293 cell line are inferior to those of non-human cell lines, such as the Chinese hamster ovary (CHO) and the murine myeloma NSO cell lines. However, the modification of genes involved in cellular processes, such as cell proliferation, apoptosis, metabolism, glycosylation, secretion, and protein folding, in addition to bioprocess, media, and vector optimization, have greatly improved the performance of this cell line. This review provides a comprehensive summary of important achievements in HEK293 cell line engineering and on the global engineering approaches and functional genomic tools that have been employed to identify relevant genes for targeted engineering.

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An anti-apoptotic HEK293 cell line provides a robust and high titer platform for transient protein expression in bioreactors

mAbs ◽

10.1080/19420862.2019.1598230 ◽

2019 ◽

Vol 11 (5) ◽

pp. 977-986 ◽

Cited By ~ 2

Author(s):

Tia A Arena ◽

Bernice Chou ◽

Peter D. Harms ◽

Athena W. Wong

Keyword(s):

Protein Expression ◽

Cell Line ◽

Hek293 Cell ◽

High Titer ◽

Hek293 Cell Line ◽

Transient Protein Expression

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Comparative Genomics, Infectivity and Cytopathogenicity of American Isolates of Zika Virus that Developed Persistent Infections in Human Embryonic Kidney (HEK293) Cells

International Journal of Molecular Sciences ◽

10.3390/ijms20123035 ◽

2019 ◽

Vol 20 (12) ◽

pp. 3035 ◽

Cited By ~ 4

Author(s):

Hebing Liu ◽

Hsiao-Mei Liao ◽

Bingjie Li ◽

Shien Tsai ◽

Guo-Chiuan Hung ◽

...

Keyword(s):

Cell Line ◽

Zika Virus ◽

Hek293 Cell ◽

Clonal Selection ◽

Gene Mutations ◽

Host Cells ◽

Hek293 Cells ◽

Specific Gene ◽

Human Embryonic Kidney ◽

Hek293 Cell Line

Zika virus (ZIKV) transmission can cause serious fetal neurological abnormalities. ZIKV persistence in various human cells and tissues can serve as infectious reservoirs and post serious threats to public health. The human embryonic kidney (HEK293) cell line with known neuronal developmental properties was readily infected by ZIKV in a strain-dependent fashion. Significant cytopathic effect in HEK293 cells infected by the prototype MR 766 strain of ZIKV resulted in complete loss of cells, while small numbers of HEK293 cells infected by contemporary ZIKV isolates (PRV or FLR strain) continued to survive and regrow to confluency in the culture around two months after initial infection. Most, if not all, of the cells in the two resulting persistently ZIKV-infected HEK293 cell lines tested positive for ZIKV antigen. Compared to HEK293 control cells, the persistently ZIKV-infected HEK293 cells had slower growth rates with some cells undergoing apoptosis in culture. The “persistent ZIKVs” produced constitutively by both PRV and FLR strains ZIKV-infected HEK293 cells had significantly attenuated cell infectivity and/or cytopathogenicity. Comparative genome sequence analyses between the persistent ZIKVs and the original inoculum ZIKVs showed no clonal selection with specific gene mutations in the prolonged process of establishing persistently PRV strain ZIKV-infected HEK293 cells; while selection of ZIKV subclones with mutations in the envelope, protein pr and multiple NS genes was evident in developing persistently FLR strain ZIKV-infected HEK293 cell line. Our study provides molecular insights into the complex interplays of ZIKV and human host cells in establishing ZIKV persistence.

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