The CD14 Knockout A2780 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human A2780 ovarian carcinoma cell line, designed to disrupt the CD14 gene encoding the lipopolysaccharide (LPS) co-receptor. This polyclonal format provides a heterogeneous loss-of-function model, avoiding the selection biases of single-cell clones while enabling robust interrogation of CD14-dependent signaling networks. The engineered cells serve as a flexible platform for studying innate immune responses, inflammation-driven cancer biology, and therapeutic resistance mechanisms in a widely used epithelial ovarian cancer background.
The A2780 parental line was established from an untreated patient with ovarian adenocarcinoma and is extensively utilized as a model for cisplatin resistance and drug sensitivity research. These adherent epithelial cells maintain key oncogenic pathways and are recognized as a standard tool for investigating ovarian tumor biology, including chemoresistance, proliferation, and metastatic behavior. Their well-characterized responsiveness to inflammatory stimuli and genotoxic stress makes them particularly suitable for examining the intersection of innate immune signaling and cancer cell phenotypes.
CD14 functions as a glycosylphosphatidylinositol-anchored co-receptor that, in concert with LPS-binding protein (LBP), captures LPS and transfers it to the TLR4/MD-2 receptor complex. This interaction triggers the recruitment of adaptor MyD88 and interleukin-1 receptor-associated kinase (IRAK), leading to activation of Lyn kinase and downstream mitogen-activated protein kinase (MAPK) cascades, including ERK, p38, and JNK. These pathways converge on transcription factors NF-??B and AP-1, driving the expression of pro-inflammatory cytokines such as TNF-??, IL-1??, and IL-6, and enzymes like inducible nitric oxide synthase (iNOS) that generate reactive oxygen species (ROS). CD14 expression is positively regulated by upstream signals including M-CSF, GM-CSF, IL-6, and the transcription factors PU.1, C/EBP, and AP-1, placing it at a critical hub of innate immune activation.
In the A2780 ovarian carcinoma context, CD14 knockout ablates the primary LPS-sensing mechanism, disrupting the LBP??CD14??TLR4/MD-2??MyD88??IRAK/TRAF6 axis. Consequently, NF-??B and MAPK signaling, along with subsequent cytokine production, are severely attenuated, creating a model that decouples cancer cells from exogenous microbial or damage-associated molecular pattern stimulation. This is especially relevant because ovarian cancer progression is influenced by inflammatory tumor microenvironments and chemoresistance mechanisms often intersect with innate immune pathways. The loss of CD14 offers a means to dissect how LPS-driven paracrine loops modulate tumor cell survival, migration, and immune cell recruitment.
Researchers can apply this knockout model in numerous advanced experimental setups, including LPS-stimulated cytokine ELISA or multiplex assays to quantify impaired TNF-??, IL-1??, and IL-6 secretion, NF-??B luciferase reporter assays to assess pathway activity, and phospho-kinase arrays to profile altered ERK, p38, and JNK phosphorylation. The cells are well-suited for co-culture experiments with macrophages or monocytes to study tumor-immune crosstalk, cisplatin sensitivity screens to evaluate the influence of CD14 on drug response, and RNA sequencing to characterize transcriptomic changes following LPS challenge. Migration, invasion, and viability assays further enable dissection of CD14-dependent metastatic traits. For additional details or customization inquiries, please contact Ascent Research.
