This product represents a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human osteosarcoma cell line 143B, targeting the androgen receptor (AR) gene. The polyclonal pool contains a heterogeneous array of cells harboring targeted disruptions within the AR locus, enabling loss-of-function analysis without isolation of individual clones. Researchers employing this model benefit from the ability to study AR-dependent phenotypes in a genetically diverse osteosarcoma background, minimizing clonal artifacts that may arise in single-cell-derived lines.
The host line, 143B, is a well-characterized human osteosarcoma cell line with established tumorigenic properties, frequently utilized in bone cancer research. Originating from an osteosarcoma patient, 143B cells exhibit metastatic potential and serve as a relevant platform for investigating molecular mechanisms underlying osteosarcoma progression, invasion, and drug response. This cell line??s adherent growth and stable karyotype facilitate reproducible experimental protocols, including transfection and pharmacological inhibition studies.
The androgen receptor is a nuclear hormone receptor and ligand-dependent transcription factor that mediates pleiotropic effects of androgens. Upon binding to dihydrotestosterone or testosterone, AR dissociates from heat shock proteins such as HSP90, translocates to the nucleus, dimerizes, and binds androgen response elements to regulate target gene expression. Key downstream targets include KLK3 (PSA), TMPRSS2, NKX3-1, and FKBP5. AR signaling integrates with multiple pathways, including PI3K/AKT, MAPK, and WNT cascades, and is modulated by coactivators like NCOA1 and NCOA2, corepressors such as NCoR, and pioneer factor FOXA1. This network controls cellular proliferation, survival, and differentiation in androgen-responsive tissues.
In the context of osteosarcoma, AR signaling may contribute to tumor growth and metastatic dissemination, although its precise role remains under investigation. The 143B osteosarcoma line, when devoid of AR function, provides a powerful tool to disentangle androgen-dependent from androgen-independent oncogenic mechanisms. This model also holds relevance for prostate cancer bone metastasis studies, as 143B cells form tumors in bone microenvironments, allowing researchers to explore AR-mediated cross-talk between tumor cells and the bone stroma. Consequently, the AR knockout polyclonal 143B cells enable interrogation of AR??s contribution to bone-tropic cancer phenotypes.
Typical experimental applications encompass biochemical, molecular, and functional assays. Western blotting and RT-qPCR can confirm loss of AR protein and transcript, along with altered expression of downstream targets like PSA and TMPRSS2. Luciferase-based ARE-reporter assays assess transcriptional activity changes. Immunofluorescence visualizes nuclear translocation defects. Proliferation (MTT, BrdU) and apoptosis (TUNEL) assays quantify growth and survival dependencies. Migration and invasion assays evaluate metastatic capacity, while drug sensitivity testing with AR antagonists such as enzalutamide reveals therapeutic vulnerabilities. For specialized inquiries, please contact Ascent Research.
