GPX8 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa human cervical adenocarcinoma cell line, engineered to disrupt the GPX8 gene. This loss-of-function model enables investigation of GPX8, an endoplasmic reticulum (ER)-resident glutathione peroxidase critical for regulating ER redox homeostasis and oxidative protein folding. The polyclonal population contains a heterogeneous spectrum of GPX8 gene disruptions, providing a robust tool for studying GPX8-dependent cellular processes without clonal selection bias.
The host HeLa cell line is an HPV18-positive human cervical epithelial cell line, originally derived from a cervical adenocarcinoma of Henrietta Lacks. These immortalized cells are widely employed as a model system for cancer biology, cell signaling, and host?Cpathogen interactions. HeLa cells exhibit robust ER stress responses and high oxidative protein folding capacity, making them an ideal platform for dissecting the functions of ER-resident redox enzymes such as GPX8.
GPX8 is an ER-resident glutathione peroxidase that reduces hydrogen peroxide and organic hydroperoxides to water and corresponding alcohols using reduced glutathione (GSH) as an electron donor, thereby maintaining ER redox balance. It directly interacts with the oxidative folding machinery: GPX8 modulates ERO1?? activity and associates with PDI, PRDX4, and the chaperone GRP78. GPX8 expression is transcriptionally upregulated by NRF2 in response to oxidative stress, and by ATF4 downstream of the PERK branch of the unfolded protein response (UPR); NF-??B and AP-1 also regulate its expression. Downstream, GPX8 attenuates ER hydrogen peroxide levels, protects against oxidative damage to proteins and lipids, and supports productive disulfide bond formation. The coupled pathway involves GSH-dependent peroxide reduction, glutathione reductase-mediated GSSG recycling, and NADPH regeneration via the pentose phosphate pathway, linking antioxidant defense to ER proteostasis.
In HeLa cells, disruption of GPX8 is anticipated to perturb ER redox homeostasis, leading to heightened sensitivity to oxidative stressors and potential activation of the UPR via IRE1??, PERK, and ATF6 sensors. Given the involvement of GPX8 in cancer progression, metastasis, and chemoresistance, this knockout model offers a physiologically relevant system to examine how loss of ER-specific antioxidant capacity influences tumor cell behavior. The HPV-positive background further allows exploration of viral-host interactions in redox regulation. Consequently, these cells are valuable for delineating the molecular mechanisms by which GPX8 contributes to malignant phenotypes and therapeutic resistance.
These GPX8 Knockout HeLa Polyclonal Cells are suitable for a broad range of experimental applications, including dissection of ER redox regulation in cancer cells, elucidation of chemoresistance mechanisms, investigation of oxidative protein folding dynamics, and evaluation of GPX8 as a therapeutic target. Researchers can employ Western blotting for GPX8 and ER stress markers (e.g., GRP78, CHOP), RT-qPCR for transcript analysis, DCFH-DA flow cytometry for ROS detection, glutathione quantification (GSH/GSSG ratios), cell viability assays under H2O2-induced stress, immunofluorescence to monitor ER localization, and functional assays such as migration, invasion, and caspase-3 activity measurements. For further information or custom inquiries, please contact Ascent Research.
