HIF‑1α and RACGAP1 promote the progression of hepatocellular carcinoma in a mutually regulatory way

Abstract

Hypoxia, a condition characterized by low oxygen levels, serves an important role in the progression of hepatocellular carcinoma (HCC). However, the precise molecular mechanisms underlying hypoxia-induced HCC progression are yet to be fully elucidated. The present study assessed the involvement of two key factors, hypoxia-inducible factor-1α (HIF-1α) and Rac GTPase activating protein 1 (RACGAP1), in HCC development under hypoxic conditions. HIF-1α and RACGAP1 genes were overexpressed and knocked down in Hep3B and Huh7 cells using lentiviral transduction and the levels of HIF-1α and RACGAP1 in the cells were assessed using quantitative PCR, western blotting and immunofluorescence. Co-immunoprecipitation experiments were performed to evaluate the interaction between HIF-1α and RACGAP1. Subsequently, the proliferation, apoptosis, migration and invasion of Hep3B and Huh7 cells were assessed using the Cell Counting Kit-8 assay, flow cytometry, Transwell assay and migration experiments. The expression levels of HIF-1α and RACGAP1 in normal and HCC tumor samples were analyzed utilizing the Gene Expression Profiling Interactive Analysis database. Furthermore, correlations between HIF-1α/RACGAP1 gene expression levels and patient survival outcomes were evaluated using the Kaplan-Meier plotter. Knockdown of HIF-1α resulted in a significant decrease in RACGAP1 expression, whilst overexpression of HIF-1α resulted in a significant increase in RACGAP1 expression. Moreover, overexpression and knockdown of RACGAP1 had the same effect on HIF-1α expression. Additionally, it was demonstrated that HIF-1α and RACGAP1 interacted directly within a complex. Overexpression of HIF-1α or RACGAP1 significantly increased proliferation, invasion and migration, and significantly decreased the proportion of apoptotic Hep3B and Huh7 cells. Conversely, knockdown of HIF-1α or RACGAP1 significantly decreased proliferation, invasion and migration, and significantly increased the proportion of apoptotic Hep3B and Huh7 cells. In addition, the combined knockdown or overexpression of HIF-1α and RACGAP1 had a more pronounced effect on HCC cell migration compared with knockdown of HIF-1α alone. Furthermore, there was a significant positive correlation between the expression levels of HIF-1α and RACGAP1 in HCC tissues and patients with HCC and upregulation of both HIF-1α and RACGAP1 demonstrated a lower overall survival probability. In conclusion, HIF-1α and RACGAP1 may synergistically contribute to the development of HCC, highlighting their potential as valuable targets for HCC therapy.

Elevated RACGAP1 Expression Enhances Malignant Potential in Lung Adenocarcinoma and Serves as a Prognostic Factor

“…For instance, it has been elucidated that RACGAP1 can enhance the aggressiveness of tumors and facilitate lymph node metastasis in colorectal cancer 6 . Additionally, in hepatocellular carcinoma, RACGAP1 interacts with HIF-1alpha, thereby exerting influence on hepatocarcinogenesis 7 . Moreover, RACGAP1 has emerged as a pivotal player in cervical cancer 8 .…”