Related Experiment Video
Updated: Jan 7, 2026
Characterization of Functionally Associated miRNAs in Glioblastoma and their Engineering into Artificial Clusters for Gene Therapy
Published on: October 4, 2019
Development of a Dual Gene-Targeted Multi-Sirna with Branched Structure and Its Role in the Therapy of Liver Cancer
Mingdong Lu1,2, Wenqin Jiang1,2, Zhekai Liu1
1Key Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, China.
Abstract:
Background: Hepatocellular carcinoma (HCC) remains a major global health challenge with limited therapeutic options. Although RNA interference (RNAi) enables precise gene silencing, its clinical application is restricted by siRNA instability, inefficient cellular uptake, and the requirement for potentially toxic delivery carriers. To address these limitations, a dual-targeted branched siRNA nanostructure (GT-multi-siRNA) was developed to simultaneously silence two HCC-related oncogenes, GP73 and hTERT. Methods: GT-multi-siRNA was synthesized in Escherichia coli and characterized for particle size, stability, Dicer processing efficiency, intracellular retention, and cytotoxicity. Its therapeutic effects were evaluated through gene-silencing assays, proliferation and migration assays in Hep3B cells, and intratumoral administration in a xenograft mouse model. Histopathology and cytokine profiling were conducted to assess biosafety. Results: GT-multi-siRNA formed uniform nanoparticles (50-100 nm) with moderate physicochemical stability and minimal cytotoxicity at concentrations ≤ 200 ng/μL. The nanostructure was efficiently processed by Dicer into functional siRNAs and remained detectable intracellularly for at least 36 h. In Hep3B cells, GT-multi-siRNA reduced GP73 and hTERT mRNA and protein levels by approximately 50%, accompanied by significant inhibition of cell proliferation and migration. In vivo, a single intratumoral dose suppressed tumor growth, while a two-dose regimen markedly limited tumor progression. No liver toxicity was observed, and cytokine analysis showed selective IL-4 upregulation without influencing IL-6 levels. Conclusions: GT-multi-siRNA demonstrates potent dual-gene silencing activity and favorable biosafety, providing a promising RNAi-based therapeutic strategy for targeted HCC treatment.
Insights
A novel dual-targeted branched siRNA nanostructure (GT-multi-siRNA) effectively silenced two oncogenes in hepatocellular carcinoma (HCC) cells. This RNA interference (RNAi) therapy showed promising results for targeted HCC treatment with good biosafety.
Area of Science:
- Biotechnology
- Nanomedicine
- Cancer Therapy
Background:
- Hepatocellular carcinoma (HCC) presents significant global health challenges with limited treatment options.
- Current RNA interference (RNAi) therapies face hurdles like siRNA instability and inefficient delivery.
- A dual-targeted nanostructure was engineered to overcome these limitations for HCC treatment.
Purpose of the Study:
- To develop and evaluate a dual-targeted branched siRNA nanostructure (GT-multi-siRNA) for simultaneous silencing of GP73 and hTERT oncogenes in HCC.
- To assess the physicochemical properties, cellular uptake, and biosafety of the GT-multi-siRNA nanostructure.
- To investigate the therapeutic efficacy of GT-multi-siRNA in vitro and in vivo models of HCC.
Main Methods:
- Synthesis and characterization of GT-multi-siRNA nanostructures in E. coli.
- Evaluation of particle size, stability, Dicer processing, intracellular retention, and cytotoxicity.
- Assessment of gene silencing, cell proliferation, and migration inhibition in Hep3B cells.
- Intratumoral administration in a xenograft mouse model, followed by histopathology and cytokine profiling.
Main Results:
- GT-multi-siRNA formed uniform nanoparticles (50-100 nm) with minimal cytotoxicity.
- Efficient Dicer processing and sustained intracellular presence (≥36 h) were observed.
- Significant reduction in GP73 and hTERT expression (approx. 50%) and inhibition of cell proliferation/migration in vitro.
- Suppressed tumor growth in vivo with a favorable safety profile, including no observed liver toxicity and selective IL-4 upregulation.
Conclusions:
- GT-multi-siRNA exhibits potent dual-gene silencing capabilities against HCC-related oncogenes.
- The nanostructure demonstrates favorable physicochemical properties and excellent biosafety.
- GT-multi-siRNA represents a promising RNAi-based therapeutic strategy for targeted hepatocellular carcinoma treatment.
More Related Videos
07:23Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells
Published on: May 30, 2025
09:18TRUE Gene Silencing: Screening of a Heptamer-type Small Guide RNA Library for Potential Cancer Therapeutic Agents
Published on: June 2, 2016
Related Concept Videos
02:57Targeted Cancer Therapies
There are several types of targeted therapies against...
02:30siRNA - Small Interfering RNAs
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...
02:15Experimental RNAi
01:22MicroRNAs
01:22MicroRNAs
00:59Gene Therapy