The identification of lncRNA SRHC as a high-priority therapeutic target underscores a significant shift toward RNA-based precision medicine in oncology. Our bioinformatics analysis reveals that while SRHC is highly conserved and robustly expressed in healthy hepatic tissue, suggesting a foundational role in maintaining liver homeostasis, it is markedly downregulated in Hepatocellular Carcinoma (HCC). This inverse correlation, corroborated by data in Table 1 and Fig. 1, implies that SRHC may function as a tumor suppressor. Mechanistically, the loss of such long non-coding RNAs often leads to derepression of oncogenic pathways, potentially through dysregulation of chromatin remodeling or the sponging of microRNAs that would otherwise inhibit proliferative signaling22,23.
The therapeutic novelty of this work is centered on the biological synergy between the restoration of this regulatory RNA and its delivery via a targeted nanoparticle system. Mechanistically, the depletion of SRHC in malignant cells likely leads to the derepression of oncogenic pathways, potentially through the loss of microRNA sponging effects or the dysregulation of chromatin remodeling complexes. By utilizing a clinically viable PLGA-based platform, we facilitate the reintroduction of SRHC to restore these disrupted homeostatic checkpoints. This approach shifts the focus from the development of novel polymer chemistries toward the strategic application of established nanotechnology to exploit the specific, previously untapped regulatory role of the SRHC axis in HCC.
Granular dispersions or solid particles with a size between 10 and 1000 nm are known as nanoparticles. Therapeutics can be delivered using nanoparticles to enhance permeability and retention (EPR). As a result, adopting nanoparticles as a delivery system for lncRNA-targeted therapy represents a multi-effect approach24. The successful conjugation of polymer nanoparticles (NPs) with SRHC yields a structurally robust, physically stable delivery system, characterized by specific physicochemical parameters that govern its behavior in biological environments. A mean particle size of 200.4 nm is generally considered optimal for systemic administration, as it is large enough to avoid rapid renal clearance yet small enough to exploit the enhanced permeability and retention (EPR) effect in targeted tissues. The Polydispersity Index (PDI) of 0.375, while bordering the upper limit for strictly monodisperse organic systems, still suggests a sufficiently homogeneous distribution for reliable drug release kinetics. Crucially, the zeta potential of 44.21 ± 0.85 mV serves as a definitive indicator of colloidal stability. In aqueous suspension, a high absolute zeta potential (> 30 mV) generates significant electrostatic repulsion between individual particles, preventing van der Waals force-driven aggregation or flocculation over time25. This high positive surface charge not only ensures a long shelf-life for the formulation but may also enhance cellular uptake through electrostatic attraction to negatively charged phospholipid bilayers on cell membranes26. The cationic surface of the nanoparticles facilitates a natural affinity for the negatively charged phospholipid bilayers of the HCC cell membranes, promoting efficient cellular internalization. Consequently, the novelty of this work is found in how these established physicochemical parameters are precisely calibrated to protect and deliver the SRHC transcript, ensuring its specific regulatory role is executed effectively within the target site.
The marginal decrease in the IC50 value for the conjugated nanoparticles (95.24 nmol vs. 98.8 nmol) suggests a subtle enhancement in bioavailability or cellular uptake facilitated by the nanoparticle carrier. SRHC inhibits cancer cell proliferation; it is highly expressed in normal liver tissue and is either absent or substantially reduced in HCC1. In a biological context, this indicates that the conjugation process does not hinder the functional activity of the lncRNA SRHC; instead, it may stabilize the RNA against enzymatic degradation or enhance its interaction with the cellular membrane, allowing for more efficient induction of cell death at lower absolute concentrations.
Rather than focusing on the development of novel polymer chemistry, the innovation of this study lies in the successful bio-functionalization of a stable delivery system to execute a specific genetic rescue mission. The data indicate that the conjugation process maintains the structural integrity and functional activity of SRHC, enabling it to effectively modulate intracellular pathways that induce apoptosis. By enhancing the bioavailability and cellular internalization of this specific lncRNA, the targeted system achieves greater therapeutic efficacy at lower absolute concentrations, establishing a strategic link between reliable nanotechnology and targeted gene regulation in HCC.
The lack of mortality and observable clinical distress suggests that both the nanoparticles (NPs) and the Slow-Release Hybrid Composite (SRHC) possess a favorable biocompatibility profile at the tested dosages. From a toxicological perspective, the absence of acute lethality and behavioral abnormalities indicates that these materials do not trigger significant systemic toxicity or neurobehavioral impairment in either physiological (normal) or pathological (diseased) models. This stability in animal welfare, characterized by the absence of stress markers such as chromodacryorrhea (bloody tears), lethargy, or ruffled fur, indicates that the administration of these agents does not exceed the Maximum Tolerated Dose (MTD). The absence of behavioral anomalies or metabolic instability suggests that the NPs-SRHC intervention does not trigger immediate systemic toxicity or acute inflammatory cascades. While these results establish a necessary safety baseline, they primarily serve as a physiological green light for deeper mechanistic exploration. Future studies should transition from these broad phenotypic observations to a more granular analysis of the intracellular signaling pathways and cytokine profiles. Understanding how these nanoparticles interact with the hepatic microenvironment at a molecular level will be essential to ensure that the lack of overt morbidity isn’t masking subtle, long-term alterations in homeostatic regulation.
HCC, a predominant histological subtype among primary liver tumors, is the sixth most prevalent malignancy worldwide and the fourth most prevalent in Egypt27. The development and progression of cancer is a complex, multi-step process driven by factors such as the aberrant expression of multiple genes, disruption of key signaling pathways and remodeling of the tumor microenvironment (TME)28. The prognosis for patients with HCC is still poor despite years of dedicated effort because of the high incidence of metastasis and recurrence29,30.
The elevation of Alpha-fetoprotein (AFP) is a classic indicator of hepatocyte regeneration and malignant transformation31. At the same time, the concomitant rise in Vascular Endothelial Growth Factor (VEGF) and Platelet-Derived Growth Factor (PDGF) reflects the highly angiogenic nature of HCC, as these factors drive the neovascularization required for tumor expansion. Furthermore, elevated levels of Tumor Necrosis Factor-alpha (TNF-α) indicate a chronic inflammatory microenvironment that promotes genomic instability32. Notably, Epithelial Cell Adhesion Molecule (EpCAM) expression suggests circulating tumor cells or a stem-like phenotype, which is often associated with higher recurrence rates and chemoresistance33. Collectively, the synergistic analysis of these biomarkers across the study groups provides a molecular snapshot of the transition from chronic liver disease to overt malignancy.
The significant reduction in tumor-associated biomarkers, specifically AFP, VEGF, TNF-α, PDGF, and EpCAM, following treatment with SRHC and conjugated nanoparticles (NPs) suggests a multi-targeted suppression of hepatocellular carcinoma (HCC) progression. The concurrent downregulation of Vascular Endothelial Growth Factor (VEGF) and Platelet-Derived Growth Factor (PDGF) indicates a disruption in the angiogenic signaling pathways necessary for tumor neovascularization and stromal support34. Furthermore, the decrease in Tumor Necrosis Factor-alpha (TNF-α) reflects a mitigation of the chronic inflammatory microenvironment that typically drives HCC proliferation35. At the same time, the reduction in Epithelial Cell Adhesion Molecule (EpCAM) suggests a potential loss of stemness and metastatic potential in the cancer cells. These findings align with previous reports36,37,38, which found that the lncRNA SRHC might hinder HCC cell proliferation while promoting its differentiation. Compared with the HCC group, in which these biomarkers remain pathologically elevated, SRHC-NP treatment restores a degree of homeostatic control, signaling a shift from a pro-tumorigenic state toward cellular recovery. The findings of this study are consistent with several studies showing that SRHC is a tumor suppressor in various cancer types39,40,41.
The marked therapeutic efficacy observed in the SRHC and conjugated nanoparticle groups relative to HCC group underscores the transformative potential of nanoformulations in clinical oncology. By using these specialized carriers, the study demonstrates significant improvements in the bioavailability and site-specific accumulation of therapeutic agents. This targeted approach effectively suppresses the biochemical indicators of malignancy, suggesting that the nano-platform provides a robust shield against systemic degradation, thereby allowing for a more potent interaction with the tumor site. While the current data clearly illustrate the disruption of tumor survival and angiogenic signaling, there remains a critical opportunity to further delineate the precise molecular crosstalk triggered by these interventions. Strengthening the mechanistic depth of these findings would involve a more granular analysis of how the NPs-lncRNA-SRHC interacts with downstream effectors within the intracellular environment. Exploring these pathways in greater detail will clarify whether the observed anti-tumor effects stem from direct gene silencing or a broader modulation of the tumor microenvironment, ultimately providing a more comprehensive understanding of the treatment’s biological impact.
The provided data in Fig. 4 illustrate a clear molecular shift associated with the progression and treatment of Hepatocellular Carcinoma (HCC), primarily driven by the dysregulation of the SENP1/β-catenin/HNF-4α axis. In the HCC group, significant elevations of SENP1 and β-catenin compared with normal controls indicate activation of oncogenic pathways. SENP1 is a deSUMOylating enzyme often implicated in stabilizing proteins that promote cell survival and proliferation. At the same time, β-catenin is a central effector of the Wnt signaling pathway, which facilitates tumor growth and epithelial-mesenchymal transition (EMT)41,43. Conversely, the drastic downregulation of HNF-4α in the HCC group reflects a loss of hepatocyte differentiation, as this transcription factor is essential for maintaining the mature liver phenotype.
The lack of a significant difference between the NPs-treated group and the HCC group indicates that the nanoparticles alone do not possess therapeutic properties for these specific gene expressions. Also, treatment with SRHC significantly attenuated the expression of SENP1 and β-catenin while partially restoring HNF-4α, suggesting its potential to inhibit tumor signaling and promote re-differentiation. Moreover, the most pronounced therapeutic effect was observed in the NPs-SRHC-treated group, which showed a significantly greater reduction in oncogenic markers and a higher restoration of HNF-4α than the SRHC-only group. This suggests that the polymer-based delivery system enhances the bioavailability and cellular uptake of SRHC, thereby enabling superior modulation of the underlying molecular pathology44,45. Therefore, the data demonstrate that nanoparticle-mediated delivery of SRHC effectively reverses the oncogenic SENP1/β-catenin signature and restores hepatocyte-protective HNF-4α expression, providing a promising strategy for targeted HCC therapy.
The improvement in the hepatic architecture suggests a positive effect of treatment on the HCC. Meanwhile, NPs-SRHC-treated group demonstrated the variable nuclear size of hepatocytes, lymphocytic infiltration, cholestasis, and regression of malignant change, with almost complete restoration of near normal lobular architecture (Fig. 5e). Overall, the microscopic examination of the liver tissue in the experimental groups provides insights into the effects of the different treatments on the HCC. The comparative efficacy of the treatments is evidenced by the degree of architectural restoration. While the SRHC-treated group showed a significant reversal of malignant features, the NPs-SRHC treated group suggests a more nuanced healing process. The presence of lymphocytic infiltration in this group is particularly notable, as it likely represents an active immune-mediated response against the remaining tumor cells46.
The profound restoration of hepatic lobular architecture observed in the NPs-SRHC-treated group underscores a critical therapeutic synergy between the lncRNA SRHC and its delivery vehicle. By leveraging a PLGA nanoparticle platform, we successfully bypassed the pharmacokinetic hurdles typically associated with RNA-based therapies, such as rapid systemic clearance and enzymatic degradation. This strategic pairing ensured that the therapeutic cargo achieved the necessary bioavailability and localized concentration required to effectively reprogram the malignant landscape. Rather than focusing on the development of novel polymer chemistries, this study highlights the high-impact utility of a clinically validated carrier to unlock the liver parenchyma’s latent regenerative potential.
Furthermore, the transition from a disorganized malignant phenotype to a structured, regenerative state provides robust evidence that the novelty of this work lies in the regulatory precision of SRHC. When delivered via an optimized nanocarrier, SRHC acts as a potent molecular switch, suppressing oncogenic signaling while simultaneously fostering a microenvironment conducive to healthy tissue repair. The consistency of the histological and biomarker data suggests that the conventional nature of the nanoparticle is a deliberate advantage; it provides a stable, predictable foundation that allows the specific, potent effects of the lncRNA to take center stage. Thus, the significance of this research resides in the successful integration of a reliable delivery system with a novel genetic target to achieve a superior healing response in hepatocellular carcinoma models.
