Panoply™ Human TFRC Over-expressing Stable Cell Line

Aberrant iron metabolism is prevalent in various tumor types, including hepatocellular carcinoma (HCC). However, the role of transferrin receptor (TFRC), a key regulator of iron metabolism involved in iron absorption, in HCC remains unclear. Here, the study shows that TFRC levels are significantly upregulated in HCC tissues compared to paired adjacent normal tissues. TFRC overexpression is positively correlated with serum alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), and tumor dysdifferentiation. Multivariate analysis indicates that TFRC upregulation is an independent predictor of poorer overall survival and disease-free survival in HCC patients. Loss of TFRC markedly impaired cell proliferation and migration in vitro and notably suppressed HCC growth and metastasis in vivo, while overexpression of TFRC performed an opposite effect. Mechanistic studies revealed that TFRC knockdown leads to downregulation of the mTOR signaling pathway, and the mTOR agonist MHY1485 can completely reverse the TFRC knockdown-induced inhibition of HCC cell biology. Furthermore, exogenous ferric citrate (FAC) or iron chelators reversed the alterations in HCC cell biological function and signaling pathway expression caused by TFRC knockdown or overexpression, respectively. These results suggest that TFRC plays an oncogenic role in HCC and may be a potential therapeutic target for inhibiting HCC progression.

To confirm the biological role of TFRC in HCC cells, TFRC expression was stably knocked down in two HCC cell lines with high TFRC expression, PLC/PRF/5 and Hep3B (PLC/PRF/5-shTFRC and Hep3B-shTFRC). Researchers also generated stable TFRC-overexpressing cells in low TFRC-expressing MHCC97H cell lines (MHCC97H-TFRC). Whether alteration of TFRC expression could affect the malignant phenotype of HCC cells was next evaluated. The proliferation potential of HCC cells was first evaluated using CCK8 and colony formation assays. The results showed that TFRC knockdown significantly reduced cell proliferation compared to the corresponding control cells (Figure 1c). Conversely, the proliferation capacity of TFRC-overexpressing cells was significantly enhanced compared to control cells (Figure 1f, g). Furthermore, Transwell migration and scratch assays indicated that downregulation of TFRC significantly inhibited cell migration compared to the corresponding control cells (Figure 1d, e), while upregulation of TFRC had the opposite effect (Figure 1h, i). In summary, these results demonstrate that TFRC is crucial for the proliferation and migration of hepatocellular carcinoma cells, confirming its oncogenic role in liver cancer.

Figure 1. TFRC aggravates HCC cell proliferation and migration in vitro. (Wang F, et al., 2024)