HnRNPA1-Regulated miR-23a-3p Promotes Hepatocellular Carcinoma Metastasis by Targeting the PTEN Signaling Pathway.

INTRODUCTION Hepatocellular carcinoma (HCC) accounts for 75-85% of primary liver cancer cases and remains a major cause of cancer-related mortality worldwide. Despite its high incidence, there is currently no definitive cure for most patients, and the underlying molecular mechanisms are not fully understood.

Methods

HCC cells were used as an in vitro HCC model. Cells were divided into five groups: control, HnRNPA1 mimic, HnRNPA1 inhibitor, miR-23a-3p mimic, and miR-23a-3p inhibitor. HnRNPA1 and miR-23a-3p were overexpressed or silenced by transient transfection with specific mimics or inhibitors, respectively. Cell migration and apoptosis were assessed using Transwell assays and Annexin V/PI-based flow cytometry. The mRNA and protein expression levels of HnRNPA1, miR-23a-3p, PTEN, and related markers were measured by quantitative real-time PCR and Western blotting. Dual luciferase reporter assays were performed to validate the direct interaction between miR-23a-3p and the 3' untranslated region of PTEN and to examine the regulatory relationship between HnRNPA1 and miR-23a-3p.

Results

Overexpression of HnRNPA1 increased its mRNA level by approximately 2.5-fold compared with the control group (P < 0.01), enhanced Hep3B cell migration by about 60%, and reduced the apoptosis rate by roughly 50% (both P < 0.01). These changes indicate that elevated HnRNPA1 expression markedly strengthens the migratory capacity of HCC cells and significantly suppresses their apoptosis, thereby conferring a clear pro-tumorigenic phenotype. Similarly, miR-23a-3p overexpression elevated its expression by about 3.3-fold (P < 0.01), promoted cell migration by approximately 57%, and decreased apoptosis by around 43% relative to control (all P < 0.01), further supporting the role of miR-23a-3p as a potent enhancer of HCC cell motility and survival. HnRNPA1 overexpression significantly up-regulated miR-23a-3p levels (≈2.1-fold, P < 0.01), suggesting that HnRNPA1 positively regulates miR-23a-3p in HCC cells. Dual-luciferase reporter assays confirmed that miR-23a-3p directly binds to the 3'-untranslated region of PTEN, and miR-23a-3p overexpression markedly decreased PTEN mRNA and protein levels by approximately 42% and 34%, respectively (P < 0.01). Functionally, restoring PTEN expression attenuated the miR-23a-3p-induced increase in cell migration by about 31% and partially reversed its anti-apoptotic effect (apoptosis increased by roughly 27% compared with the miR-23a-3p mimic group; P < 0.05), indicating that suppression of PTEN is a critical downstream event by which the HnRNPA1/miR-23a-3p axis drives HCC progression.

Discussion

The research results show that HnRNPA1, miR-23a-3p, and PTEN play crucial roles in the progression of liver cancer. These molecules interact within a regulatory network that may become a therapeutic target.

Conclusion

HnRNPA1, miR-23a-3p, and PTEN are key regulators of HCC progression and may serve as therapeutic targets for the treatment of HCC.