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Reduction sensitive CC9-PEG-SSBPEI/miR-148b nanoparticles: Synthesis, characterization, targeting delivery and
Cui Cheng1, Zhihong Zhang1, Siyuan Wang1
1Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou, 350002, China.
Abstract:
miRNAs such as miR-148b play crucial regulatory role in tumor metastasis, but their applications are limited because they are easy to degrade in serum conditions and lack targeting ability. Herein, CC9-PEG-SSBPEI was synthesized and used as nano-carrier for miR-148b. DLS and gel retardation analyses indicated that CC9-PEG-SSBPEI could combine with miR-148b by charge interaction and formed into nanoparticles with the size changed from 811.6 nm to 146.4 nm. CC9-PEG-SSBPEI could protect miR-148b from RNase A degradation and showed a reduction sensitive release of miR-148b. FACS analysis and CLSM images displayed that the conjugated CC9 peptide improved the accumulation and penetration of the nanoparticles in HuH-7 liver cancer cells through binding with the target of miR-148b neuropilin-1(NRP-1) on the cell surface. The raised level of miR-148b in turn inhibited the expression of NRP-1 and suppressed the migration of HuH-7 liver cancer cells. Moreover, hemolysis and cytotoxicity assay demonstrated that the nanoparticles had good hemo- and cyto- compatibility. Hence, CC9-PEG-SSBPEI/miR-148b nanoparticles had the potential for targeting delivery of miR-148b and anti-metastasis of hepatocellular carcinoma (HCC) cells.
Insights
This study developed novel nanoparticles for miR-148b delivery, enhancing its stability and targeting ability to inhibit hepatocellular carcinoma (HCC) metastasis. The CC9-PEG-SSBPEI/miR-148b nanoparticles show promise for cancer therapy.
Area of Science:
- Biomaterials Science
- Nanotechnology
- Molecular Biology
- Oncology
Background:
- MicroRNAs (miRNAs), such as miR-148b, are critical regulators of tumor metastasis.
- Current applications of miRNAs are hindered by their susceptibility to degradation and lack of targeted delivery.
- Hepatocellular carcinoma (HCC) metastasis remains a significant clinical challenge.
Purpose of the Study:
- To synthesize and characterize a novel nano-carrier (CC9-PEG-SSBPEI) for miR-148b delivery.
- To evaluate the targeting efficiency and anti-metastatic potential of the CC9-PEG-SSBPEI/miR-148b nanoparticles in liver cancer cells.
- To assess the biocompatibility of the developed nanocarrier system.
Main Methods:
- Synthesis of CC9-PEG-SSBPEI nano-carrier.
- Characterization of CC9-PEG-SSBPEI/miR-148b nanoparticles using Dynamic Light Scattering (DLS) and gel retardation assays.
- Evaluation of miR-148b protection from degradation (RNase A assay) and release kinetics.
- Assessment of cellular uptake, penetration, and targeting via Flow Cytometry (FACS) and Confocal Laser Scanning Microscopy (CLSM) in HuH-7 cells.
- Investigation of anti-metastatic effects by measuring neuropilin-1 (NRP-1) expression and cell migration.
- Hemolysis and cytotoxicity assays for biocompatibility assessment.
Main Results:
- CC9-PEG-SSBPEI successfully formed stable nanoparticles with miR-148b, reducing particle size from 811.6 nm to 146.4 nm.
- The nanoparticles protected miR-148b from degradation and exhibited reduction-sensitive release.
- The CC9 peptide facilitated enhanced cellular accumulation and penetration in HuH-7 cells by targeting cell surface NRP-1.
- Upregulated miR-148b effectively inhibited NRP-1 expression and suppressed liver cancer cell migration.
- The nanoparticles demonstrated good hemo- and cyto-compatibility.
Conclusions:
- CC9-PEG-SSBPEI serves as an effective nano-carrier for miR-148b, enhancing its stability and targeting capabilities.
- The developed CC9-PEG-SSBPEI/miR-148b nanoparticles show significant potential for targeted delivery of miR-148b.
- This system holds promise as an anti-metastatic therapeutic strategy for hepatocellular carcinoma (HCC).
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