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[Effect of siRNA silenced human tissue factor gene on hepatocellular carcinoma cell invasion and metastasis]
Qi Zhou1, Xiang-Bing Zhou, Ya-Feng Wang
1Department of Hepatobiliary Surgery, First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China.
Objective:
To study the effect of tissue factor (TF) on human hepatocellular carcinoma cells when their inner TF gene was knocked down by RNA interference.
Methods:
The eukaryotic expression vector bearing siRNA sequence that targeted at TF gene was transfected into human hepatocellular carcinoma cell line HepG2. RT-PCR and Western blot were used to detect the changes of TF gene expression. Transwell invasion assay invitro were used to show the invasive ability of HepG2 cells with transfected pGPU6/GFP/Neo-TFsiRNA.
Results:
RT-PCR and Western blot analysis showed that HepG2 cells with pGPU6/GFP/Neo-TFsiRNA transfected decreased the endogenous TF gene expression. Correspondingly the mean invading cells was 14 ± 10 for pGPU6/GFP/Neo-TFsiRNA transfection and 128 ± 18 for the NCsiRNA transfection, respectively by the Transwell invasion test in vitro (P < 0.05). The results indicated that the invasive ability of the HepG2 cells with pGPU6/GFP/Neo-TFsiRNA transfected decreased obviously.
Conclusions:
TF is involved in the progression of hepatocellular carcinoma and inhibiting the expression of TF can decrease the invasion and metastasis ability of tumor cell in vitro.
Insights
Knocking down tissue factor (TF) in hepatocellular carcinoma cells significantly reduced their invasion capabilities. This suggests TF plays a key role in liver cancer progression and metastasis.
Area of Science:
- Oncology
- Molecular Biology
Background:
- Hepatocellular carcinoma (HCC) is a major global health concern.
- Tissue Factor (TF) is implicated in cancer progression, but its specific role in HCC invasion requires further elucidation.
Purpose of the Study:
- To investigate the functional role of Tissue Factor (TF) in human hepatocellular carcinoma (HCC) cell invasion.
- To determine the effect of TF gene knockdown on HCC cell invasive potential.
Main Methods:
- Human HCC cell line (HepG2) was transfected with a eukaryotic expression vector containing siRNA targeting the TF gene.
- TF gene expression levels were assessed using RT-PCR and Western blot.
- In vitro cell invasion was evaluated using the Transwell invasion assay.
Main Results:
- Transfection with pGPU6/GFP/Neo-TFsiRNA led to a significant decrease in endogenous TF gene expression in HepG2 cells.
- The invasive ability of HepG2 cells was markedly reduced following TF gene knockdown (14 ± 10 invading cells vs. 128 ± 18 in controls, P < 0.05).
Conclusions:
- Tissue Factor (TF) is involved in the progression of hepatocellular carcinoma.
- Inhibiting TF expression effectively decreases the invasion and metastasis capabilities of HCC cells in vitro.
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