The SQSTM1 Knockout SK-OV-3 Cell Line is a CRISPR/Cas9-edited knockout cell line engineered to disrupt the SQSTM1 gene in the human ovarian adenocarcinoma cell line SK-OV-3. This cell-based model enables loss-of-function studies of the autophagy receptor and signaling scaffold p62/SQSTM1 in a cellular background relevant to high-grade serous ovarian carcinoma. The knockout cell line is provided as a stable cell population suitable for downstream functional assays, offering researchers a defined tool to interrogate SQSTM1-dependent mechanisms without the variability of transient knockdown approaches. The gene disruption is achieved through CRISPR/Cas9-mediated targeting, resulting in a robust knockout model for applications in cancer biology, autophagy research, and drug resistance studies.
The host cell line SK-OV-3 was originally derived from the ascites of a 64-year-old female patient with ovarian adenocarcinoma and exhibits a near-triploid karyotype characteristic of aggressive epithelial tumors. This cell line is extensively validated as a model for high-grade serous ovarian carcinoma, displaying relevant oncogenic mutations and signaling pathway alterations. SK-OV-3 cells are widely used in ovarian cancer research to investigate tumorigenesis, metastasis, and response to chemotherapeutic agents such as cisplatin. The epithelial nature of these cells provides a physiologically appropriate context for studying autophagy-mediated processes that influence ovarian cancer progression and treatment outcomes.
SQSTM1 encodes the multifunctional protein p62, which serves as a critical autophagy receptor by binding ubiquitinated cargo through its UBA domain and tethering it to autophagosomal membranes via interaction with LC3 and GABARAP family members. Beyond its role in selective autophagy, p62 functions as a signaling scaffold that integrates multiple stress-responsive pathways. It is transcriptionally regulated by NRF2 and NF-??B and, in turn, modulates mTORC1 activity by interacting with Raptor and TRAF6. Under oxidative stress, p62 is phosphorylated by protein kinase C and AMPK, which influences its oligomerization and receptor function. The protein forms complexes with Keap1, leading to NRF2 stabilization and antioxidant response activation, while also interacting with MEKK3 and PKC?? to regulate NF-??B and MAPK/ERK signaling cascades. Consequently, SQSTM1 knockout disrupts autophagy flux and perturbs the crosstalk between mTOR, NRF2, and NF-??B pathways.
Disruption of SQSTM1 in SK-OV-3 cells provides a powerful system to dissect the contribution of p62 to ovarian cancer pathophysiology. Impaired autophagy flux resulting from p62 loss leads to accumulation of damaged mitochondria and protein aggregates, potentially sensitizing cells to metabolic stress and altering apoptotic thresholds. Because SQSTM1-dependent signaling influences chemoresistance mechanisms, this knockout model is particularly valuable for investigating how autophagy modulates responses to platinum-based therapies. Additionally, the interplay between p62 and oncogenic pathways such as mTOR and NF-??B can be systematically examined in this epithelial tumor context, offering insights into tumor growth regulation and metastatic potential. The model thus bridges basic autophagy biology and translational ovarian cancer research.
Researchers can employ the SQSTM1 Knockout SK-OV-3 Cell Line in a variety of experimental workflows. Western blotting for LC3 lipidation and p62 accumulation, combined with autophagy flux assays using bafilomycin A1, enables quantitative assessment of autophagy defects. Immunofluorescence microscopy for LC3 puncta provides spatial resolution of autophagosome formation. Functional studies may include cell viability assays with cisplatin to evaluate chemosensitivity, RT-qPCR profiling of autophagy-related genes, and flow cytometry-based apoptosis detection. Migration and invasion assays further extend the utility to metastasis research. This knockout cell line is ideally suited for mechanistic studies, drug screening for autophagy modulators, and validation of p62 as a therapeutic target. For technical specifications and ordering information, please contact Ascent Research.
