PPP5C Knockout HCT 116 Cell Line

The PPP5C Knockout HCT 116 Cell Line is a CRISPR/Cas9-edited knockout cell line targeting the PPP5C gene in the human HCT 116 colorectal carcinoma background. This loss-of-function model disrupts PPP5C expression, enabling investigation of serine/threonine protein phosphatase 5 in cellular signaling and cancer biology.

HCT 116 cells were derived from a male colorectal carcinoma patient and are widely employed in cancer research and drug screening due to their distinct genetic features, including microsatellite instability (MSI-H), MLH1 deficiency, and mutations in KRAS (G13D) and CTNNB1. These alterations drive constitutive MAPK and Wnt pathway activity, and the near-diploid karyotype provides a stable genomic background for knockout studies.

PPP5C encodes the catalytic subunit of protein phosphatase 5 (PP5), a serine/threonine phosphatase that dephosphorylates key signaling molecules such as MAP3K5 (ASK1), ATM, ATR, and the glucocorticoid receptor (NR3C1). PP5 is regulated by HSP90 (HSP90AA1/HSP90AB1) and the co-chaperone CDC37, and its activity is modulated by reactive oxygen species and arachidonic acid. Through dephosphorylation of ASK1, PP5 suppresses stress-induced JNK and p38 MAPK cascades, while dephosphorylation of ATM and ATR attenuates DNA damage checkpoint signaling. Dysregulation of PP5 is associated with multiple cancers, including colorectal, breast, and hepatocellular carcinomas, where it contributes to impaired apoptosis and aberrant cell cycle progression.

In the HCT 116 context, PPP5C knockout is particularly relevant for colorectal cancer research. The MSI-H status and KRAS mutation create a dependency on DNA repair and MAPK pathways, and PP5??s role in modulating ATM/ATR signaling suggests that its loss may sensitize cells to genotoxic stress. Additionally, the CTNNB1 mutation activates Wnt signaling, and cross-talk with the Hippo pathway through PP5 may impact tumor cell growth. This model thus facilitates the study of PPP5C in signaling networks commonly perturbed in colorectal tumors.

The PPP5C Knockout HCT 116 Cell Line supports diverse experimental approaches, including Western blotting and phospho-signaling analysis to monitor pathway activation, immunofluorescence and flow cytometry for subcellular localization and cell cycle profiling, and apoptosis or colony formation assays to evaluate therapeutic responses. It is ideal for drug target validation, screening of PPP5C inhibitors, and co-immunoprecipitation studies of PP5-containing complexes. Applications also extend to glucocorticoid receptor signaling and DNA damage repair. For further information, contact Ascent Research.