The CASP3 Knockout HGC-27 Polyclonal Cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population originating from the HGC-27 human gastric carcinoma epithelial cell line. This product provides a heterogeneous mixture of cells carrying targeted disruption of the CASP3 locus, resulting in loss of caspase-3 protein expression and function. Unlike monoclonal knockout lines, the polyclonal format preserves the genetic diversity inherent in the parental cell pool while ensuring effective knockout at the population level. The model is designed for advanced functional studies of caspase-3-dependent apoptosis in a metastatic gastric cancer background.
The HGC-27 cell line was established from the lymph node metastasis of a human gastric adenocarcinoma, and it exhibits epithelial morphology characteristic of gastric carcinoma cells. As a widely used model for gastric cancer research, HGC-27 retains key features of metastatic disease, including aggressive growth and altered signaling networks. The cellular background provides a clinically relevant context to explore how loss of caspase-3 function influences apoptotic and survival pathways in a tumor type often resistant to conventional therapies. CASP3 disruption in this setting enables precise dissection of apoptosis evasion mechanisms.
CASP3 encodes caspase-3, a critical executioner caspase that mediates the terminal events of apoptotic cell death. Upon activation by initiator caspases such as CASP8, CASP9, and CASP10 via extrinsic or intrinsic pathways, or by granzyme B during immune-mediated killing, caspase-3 proteolytically cleaves a diverse set of downstream substrates, including PARP1, ICAD (DNA fragmentation factor), lamin A/C, gelsolin, ROCK1, and PKC??. This cleavage leads to hallmark apoptotic features: DNA fragmentation, nuclear condensation, and membrane blebbing. Caspase-3 activity is tightly regulated by inhibitor of apoptosis proteins like XIAP, which is antagonized by SMAC/DIABLO released from mitochondria. Upstream signaling involves death receptor ligand systems such as FasL/Fas and TNF??/TNFR1, the formation of the death-inducing signaling complex (DISC), and mitochondrial events driven by Bcl-2 family members, cytochrome c, and APAF1 within the apoptosome. Thus, CASP3 sits at a convergence point of multiple apoptotic pathways.
In HGC-27 gastric carcinoma cells, CASP3 knockout serves as a powerful tool to investigate the molecular determinants of apoptosis resistance, a hallmark of cancer. Gastric cancers frequently exhibit dysregulated apoptotic signaling, and the loss of caspase-3 function in this metastatic derivative allows researchers to study how tumor cells evade cell death and develop chemoresistance. By comparing knockout and wild-type cells, key nodes for therapeutic intervention can be identified, particularly those involving upstream regulators like CASP8 or XIAP. Moreover, this model facilitates exploration of caspase-3-independent cell death pathways that may be exploited in drug-resistant gastric malignancies.
Researchers can employ the CASP3 Knockout HGC-27 Polyclonal Cells in a variety of experimental workflows, including western blot analysis for cleaved caspase-3, caspase-3 activity assays using fluorogenic substrates, and flow cytometric detection with FITC-DEVD-FMK to monitor active caspase-3. The model is suited for apoptosis induction studies with chemotherapeutic drugs or apoptotic stimuli, followed by Annexin V/PI staining or TUNEL assays to quantify cell death. Additionally, PARP cleavage assays provide a reliable readout of caspase-3-dependent events. These applications support drug screening for pro-apoptotic compounds, functional mapping of caspase-3 substrates, and mechanistic studies of gastric cancer biology. For further information or assistance, please contact Ascent Research.
