Related Experiment Video
Updated: Jun 11, 2026
Stem Cell-Derived Viral Ag-Specific T Lymphocytes Suppress HBV Replication in Mice
Published on: September 25, 2019
Suppression of human hepatoma (HepG2) cell growth by nuclear factor-kappaB/p65 specific siRNA
Wei Wu1, Dengfu Yao, Yilang Wang
1Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, 20 West Temple Road, Nantong 226001, Jiangsu Province, China.
Abstract:
Nuclear factor-kappaB (NF-κB) is a transcription factor and antagonist of apoptosis during liver regeneration and closely related to the formation and development of hepatocellular carcinoma. In the present study, we investigated the effect of small interference RNA (siRNA)-mediated inhibition of NF-κB on growth of human hepatoma (HepG2) cells. Our data indicated that the expression of NF-κB/p65 mRNA was significantly higher in the HepG2 cells than that in the normal liver (LO2) cells before transfection, and the expression of NF-κB/p65 in the HepG2 cells with NF-κB/p65 siRNA (100 nMol/L) transfection at 72 h was reduced at the levels of mRNA (93%) and protein (62%) using real-time reverse transcription-polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blotting. Interestingly, the apoptosis index of the HepG2 cells increased up to 85%, detected by Annexin V-fluorescein isothiocyanate, suggesting that NF-κB is overexpressed in hepatoma cells and can be inhibited by NF-κB/p65 siRNA through the apoptotic mechanism. Thus, we conclude that NF-κB is a potential molecular target for HCC gene therapy.
Insights
Nuclear factor-kappaB (NF-κB) is overexpressed in liver cancer cells. Inhibiting NF-κB with siRNA significantly reduced cancer cell growth and increased apoptosis, suggesting its potential as a gene therapy target for hepatocellular carcinoma (HCC).
Area of Science:
- Molecular Biology
- Oncology
- Hepatology
Background:
- Nuclear factor-kappaB (NF-κB) plays a crucial role in liver regeneration and hepatocellular carcinoma (HCC) development.
- NF-κB acts as an antagonist of apoptosis, contributing to cancer progression.
Purpose of the Study:
- To investigate the effect of inhibiting NF-κB using small interference RNA (siRNA) on human hepatoma (HepG2) cell growth.
- To explore NF-κB as a potential molecular target for HCC gene therapy.
Main Methods:
- Real-time reverse transcription-polymerase chain reaction (RT-PCR) to measure mRNA levels.
- Enzyme-linked immunosorbent assay (ELISA) and Western blotting to assess protein expression.
- Annexin V-fluorescein isothiocyanate (FITC) staining to detect apoptosis.
Main Results:
- NF-κB/p65 mRNA expression was significantly higher in HepG2 cells compared to normal LO2 cells.
- NF-κB/p65 siRNA transfection reduced mRNA by 93% and protein by 62% in HepG2 cells.
- Apoptosis index in HepG2 cells increased by 85% after siRNA transfection.
Conclusions:
- NF-κB is overexpressed in hepatoma cells.
- NF-κB/p65 siRNA effectively inhibits NF-κB expression, inducing apoptosis in HepG2 cells.
- NF-κB represents a promising molecular target for HCC gene therapy.
Related Concept Videos
02:23Abnormal Proliferation
02:26NF-κB-dependent Signaling Pathway
NF-κB-dependent Signaling Mechanism
The heterodimer of NF-κB...
01:12Loss of Tumor Suppressor Gene Functions
When the tumor suppressor genes develop mutations or are lost, cells start growing out of control, leading to cancer. However, a single functional copy of the tumor suppressor gene is enough for the cells to maintain their normal functions and cell...