Panoply™ Human CTGF Knockdown Stable Cell Line

Since its introduction in 2008, sorafenib has served as the standard first-line systemic therapy for advanced hepatocellular carcinoma (HCC). However, the rapid emergence of drug resistance often limits its clinical efficacy. Here, researchers investigated the molecular mechanisms underlying sorafenib resistance, focusing on the role of connective tissue growth factor (CCN2/CTGF) in regulating the c-Met signaling pathway. The results demonstrated that CCN2 overexpression not only significantly promoted HCC cell proliferation but also induced resistance to sorafenib. Mechanistic studies revealed that CCN2 binds to integrin αV, triggering focal adhesion kinase (FAK) phosphorylation; this subsequently promotes the nuclear translocation of Yes-associated protein (YAP), ultimately leading to the transcriptional upregulation of c-Met. The existence of this signaling axis was confirmed through tyrosine kinase PCR arrays, co-immunoprecipitation, and Western blot analysis. In vivo experiments further demonstrated that simultaneously targeting CCN2 in combination with sorafenib produced a synergistic effect, significantly inhibiting tumor growth and restoring drug sensitivity. These findings not only reveal a novel CCN2/FAK/YAP/c-Met signaling axis involved in sorafenib resistance but also provide a mechanistic basis for dual-targeting strategies to improve therapeutic outcomes in advanced HCC.

Immunohistochemical analysis revealed that CCN2 was primarily localized in the cytoplasm of hepatocellular carcinoma (HCC) cells (Figure 1A). CCN2 expression was significantly correlated with a history of hepatitis B, aspartate aminotransferase (AST) levels, the severity of liver cirrhosis, and tumor size. Consistent with clinical findings, Western blot analysis demonstrated differential CCN2 protein expression across various HCC cell lines; specifically, CCN2 expression was significantly elevated in Hep3B and Huh-7 cell lines compared to the normal liver cell line LO2 (Figure 1B). To investigate the functional role of CCN2, researchers generated stable CCN2-overexpressing LM3 cells and CCN2-knockdown Hep3B/Huh-7 cells (Figures 1C and 1D). CCK-8 and colony formation assays indicated that cell proliferation was significantly inhibited in CCN2-knockdown cells but promoted in CCN2-overexpressing cells (Figures 1E and F). Similarly, Transwell migration assays showed that CCN2 depletion inhibited cell migration (Figure 1G), whereas wound healing assays confirmed that CCN2 overexpression enhanced migratory capacity (Figure 1H).

Figure 1. Expression and biological functions of CCN2 in HCC. (Cui L, et al., 2025)