Mechanisms of CRLF3-targeted binding to ACTR2 to promote hepatocellular carcinoma progression and effects on the
SuSu Ye1, XinLei Zhang2, FengChao Liu1
1Liver Disease Center, The Affiliated Hospital of Qingdao University, No. 59 Haier Road, Laoshan District, Qingdao City, 266035 Shandong Province China.
Abstract:
This study aimed to delve deeper into the effects of CRLF3 on the immune microenvironment and the interaction between CRLF3 and ACTR2 in hepatocellular carcinoma (HCC). CRLF3 and ACTR2 in mouse tumor tissues and HepG2 cells were measured by RT-qPCR and Western Blot. The proliferative ability of HepG2 cells was assessed by MTT and colony formation assays, with apoptosis determined by flow cytometry, and migration and invasion quantified by Transwell assay. The apoptosis rate of CD8+ T cells was calculated by flow cytometry, as well as TNF-α and IFN-γ positivity in CD8+ T cells. TNF-α, IFN-γ, and IL-2 were assayed by ELISA. The interaction between CRLF3 and ACTR2 was examined using immunoprecipitation and Western Blot experiments. CRLF3 targeted binding to ACTR2 promoted the proliferative and migratory capacities of HepG2 cells and inhibited apoptosis. Lowering CRLF3 inhibited HCC cell immune escape, with a significant increase in TNF-α and IFN-γ-positive populations in CD8+ T cells, and enhancing ACTR2 significantly mitigated this effect. Lowering CRLF3 inhibited HCC xenografted tumor growth in nude mice. Through its targeted binding to ACTR2, CRLF3 aids in the growth and immune escape of HCC cells.
Insights
This study reveals that Cytokine Receptor Like Factor 3 (CRLF3) promotes hepatocellular carcinoma (HCC) growth and immune evasion by binding to ACTR2. Inhibiting CRLF3 reduces tumor growth and enhances anti-tumor immunity in HCC.
Area of Science:
- Oncology
- Immunology
- Molecular Biology
Background:
- Hepatocellular carcinoma (HCC) remains a significant global health challenge.
- Understanding the molecular mechanisms driving HCC progression and immune evasion is crucial for developing effective therapies.
Purpose of the Study:
- To investigate the role of Cytokine Receptor Like Factor 3 (CRLF3) in hepatocellular carcinoma (HCC).
- To elucidate the interaction between CRLF3 and ACTR2 in the context of HCC.
- To assess the impact of CRLF3 on the tumor immune microenvironment and HCC cell behavior.
Main Methods:
- Gene expression and protein levels of CRLF3 and ACTR2 were quantified using RT-qPCR and Western Blot.
- Cell proliferation, apoptosis, migration, and invasion were assessed using MTT, colony formation, flow cytometry, and Transwell assays.
- Immune cell responses, including CD8+ T cell apoptosis and cytokine production (TNF-α, IFN-γ, IL-2), were analyzed via flow cytometry and ELISA.
- Protein-protein interactions were confirmed using immunoprecipitation and Western Blot.
Main Results:
- CRLF3 directly binds to ACTR2, enhancing HCC cell proliferation and migration while inhibiting apoptosis.
- Reducing CRLF3 levels suppressed HCC immune escape, increasing TNF-α and IFN-γ positive CD8+ T cells.
- Enhancing ACTR2 expression partially reversed the anti-tumor effects of lowering CRLF3.
- CRLF3 inhibition led to decreased tumor growth in a mouse xenograft model.
Conclusions:
- CRLF3, through its interaction with ACTR2, promotes HCC cell proliferation, migration, and immune evasion.
- Targeting CRLF3 represents a potential therapeutic strategy to enhance anti-tumor immunity and inhibit HCC progression.
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