The GORASP2 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human hepatocellular carcinoma line, featuring disrupted GORASP2 gene expression and loss of GRASP55 protein function. This polyclonal pool preserves the genetic heterogeneity of the original line while providing a robust loss-of-function model for studying GRASP55-dependent cellular processes without requiring clonal isolation, making it suitable for diverse functional assays in cancer cell biology.
The host SK-HEP-1 cell line is a malignant liver epithelial model established from the ascites of a patient with liver adenocarcinoma. These cells exhibit key characteristics of hepatocellular carcinoma, including responsiveness to growth factors EGF and PDGF, and active ERK1/2 and mTOR signaling pathways. They are widely employed to investigate hepatocarcinogenesis, metastatic progression, and drug resistance, providing a clinically relevant background for dissecting Golgi-associated protein functions in liver cancer.
GRASP55, encoded by GORASP2, is a peripheral Golgi protein that mediates cisternal stacking and unconventional secretion of factors such as TGF-??1. It is phosphorylated by ERK1/2 and JNK downstream of EGF and PDGF receptors, and is regulated by mTOR kinase. GRASP55 interacts with GRASP65 (GORASP1), GM130 (GOLGA2), BECN1, and SEC16A, and promotes integrin ??5/??1 trafficking, LC3 lipidation during autophagy, and paxillin phosphorylation. Disruption of GORASP2 impairs Golgi stacking, reduces unconventional TGF-??1 secretion, alters integrin trafficking, and attenuates autophagosome formation, ultimately diminishing hepatocellular carcinoma cell invasion and metastasis.
In SK-HEP-1 cells, GORASP2 knockout uncouples key tumorigenic processes driven by GRASP55. The model enables dissection of Golgi architecture, mTOR/ERK signaling crosstalk, and autophagic flux in the context of liver cancer. Loss of GRASP55-dependent TGF-??1 secretion and integrin-mediated adhesion highlights its role in epithelial?Cmesenchymal transition and metastatic spread, offering insights into mechanisms of hepatocellular carcinoma progression and potential therapeutic vulnerabilities.
Research applications include hepatocellular carcinoma metastasis studies, Golgi structural biology, unconventional secretion mechanism analysis, autophagy regulation investigation, and drug resistance screening. Representative assays comprise Western blotting for GRASP55 and Golgi markers, immunofluorescence for Golgi morphology, transwell invasion, LC3 puncta analysis, ELISA for secreted TGF-??1, co-immunoprecipitation of GRASP55 interactors, and phospho-ERK signaling profiling. For further information and technical support, please contact Ascent Research.
