HnRNPA1-Regulated miR-23a-3p Promotes Hepatocellular Carcinoma Metastasis by Targeting the PTEN Signaling Pathway
Weiwei Wang, Sen Lu, Feng Jiao, Dayong Zhou, Guoyong Han, Jinwei Qi, Xinli HuangIntroduction:
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.