The HSPA4 Knockout HCCLM3 Cell Line is a CRISPR/Cas9-edited human liver cancer model featuring targeted disruption of the HSPA4 gene (encoding an HSP70 family chaperone) in the HCCLM3 hepatocellular carcinoma cell background. This cell line provides a stable loss-of-function system for investigating HSP70-mediated processes without pharmacological inhibition. The knockout is achieved using CRISPR/Cas9 technology to eliminate functional HSPA4 protein expression.
HCCLM3 is a highly metastatic variant derived from the MHCC97H cell line, originally established from a Chinese patient with HBV-related hepatocellular carcinoma. This cell line is characterized by strong invasive and migratory capacity, making it a well-suited model for metastasis research. HCCLM3 cells harbor mutations in TP53 and other oncogenic pathways, providing a relevant genetic background for studying liver cancer progression.
HSPA4 encodes a molecular chaperone central to protein folding, stress responses, and aggregate clearance. Transcriptionally activated by HSF1 in response to hyperthermia, oxidative stress, or heavy metals, HSPA4 cooperates with co-chaperones such as HOP, HSP40/DNAJ, BAG proteins, and the ubiquitin ligase CHIP/STUB1, and works alongside HSP90 to fold client proteins including kinases and steroid receptors. It also contributes to ER-associated degradation and antigen processing pathways. Disruption of HSPA4 impairs proteostasis and sensitizes cells to proteotoxic stress.
In the HCCLM3 background, loss of HSPA4 is anticipated to disrupt protein homeostasis, particularly under tumor microenvironmental stress. This model enables dissection of how chaperone dysfunction affects hepatocellular carcinoma behaviors such as proliferation, migration, and survival. Additionally, because HSPA4 interfaces with the MAPK cascade (RAS-RAF-MEK-ERK), the knockout may alter oncogenic signaling, making it a valuable tool for studying crosstalk between stress pathways and metastasis.
Typical applications include cell viability assays (MTT/CCK-8) under stress conditions, Transwell migration/invasion assays, Annexin V apoptosis detection, and co-immunoprecipitation to examine altered interactomes of HSP70 partners. Transcriptomic analysis via RNA-seq can reveal global expression changes. The cell line also supports targeted inhibitor testing against heat shock or MAPK components. For technical inquiries, please contact Ascent Research.
