Related Experiment Video
Updated: Jul 8, 2026
TRUE Gene Silencing: Screening of a Heptamer-type Small Guide RNA Library for Potential Cancer Therapeutic Agents
Published on: June 2, 2016
Gli-1 siRNA induced apoptosis in Huh7 cells
Xi-Lin Chen1, Liang-Qi Cao, Miao-Rong She
1Department of Hepatobiliary Surgery, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510080, Guangdong Province, China.
Aim:
To investigate the effects of Gli-1 small interference RNA (siRNA) on Huh7 cells, and the change of Bcl-2 expression in Huh7 cells.
Methods:
Human hepatocellular carcinoma cells Huh7 were used. Cell viability was analyzed by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. The expressions of Gli-1 and Bcl-2 family members were detected by RT-PCR and Western blot. Apoptosis was detected by Flow cytometry using propidium iodide, measured by Hoechst 33258 staining using Advanced Fluorescence Microscopy and caspase-3 enzymatic assay. Cell growth was analyzed after treatment with Gli-1 siRNA and 5-fluorouracil (5-Fu).
Results:
Inhibition of Gli-1 mRNA in Huh7 cells through Gli-1 siRNA reduced cell viability. Gli-1 siRNA treatment also induced apoptosis by three criteria, increase in the sub-G1 cell cycle fraction, nuclear condensation, a morphologic change typical of apoptosis, and activation of caspase-3. Gli-1 siRNA was also able to down-regulate Bcl-2. However, Gli-1 siRNA resulted in no significant changes in Bcl-xl, Bax, Bad, and Bid. Furthermore, Gli-1 siRNA increased the cytotoxic effect of 5-Fu on Huh7 cell.
Conclusion:
Down-regulation of Bcl-2 plays an important role in apoptosis induced by Gli-1 siRNA in HCC cells. Combination Gli-1 siRNA with chemotherapeutic drug could represent a more promising strategy against HCC. The effects of the strategies need further investigation in vivo and may have potential clinical application.
Insights
Gli-1 small interference RNA (siRNA) inhibits hepatocellular carcinoma cell viability and induces apoptosis by down-regulating Bcl-2. Combining Gli-1 siRNA with 5-fluorouracil enhances its cytotoxic effect on HCC cells.
Area of Science:
- Hepatocellular Carcinoma Research
- Molecular Biology
- Cancer Therapeutics
Background:
- Hepatocellular carcinoma (HCC) remains a significant global health challenge.
- The Hedgehog signaling pathway, particularly Gli-1, is implicated in HCC progression.
- Targeting Gli-1 offers a potential therapeutic strategy for HCC.
Purpose of the Study:
- To investigate the effects of Gli-1 small interference RNA (siRNA) on Huh7 HCC cells.
- To analyze the impact of Gli-1 siRNA on Bcl-2 expression and apoptosis.
- To evaluate the combined effect of Gli-1 siRNA and 5-fluorouracil (5-Fu) on HCC cells.
Main Methods:
- Huh7 human HCC cells were treated with Gli-1 siRNA.
- Cell viability assessed using MTT assay.
- Gene and protein expression analyzed by RT-PCR and Western blot.
- Apoptosis evaluated via flow cytometry, Hoechst staining, and caspase-3 assay.
- Combined treatment with Gli-1 siRNA and 5-Fu was assessed.
Main Results:
- Gli-1 siRNA significantly reduced Huh7 cell viability.
- Gli-1 siRNA induced apoptosis, evidenced by increased sub-G1 fraction, nuclear condensation, and caspase-3 activation.
- Gli-1 siRNA down-regulated Bcl-2 expression but did not affect Bcl-xl, Bax, Bad, or Bid.
- Co-treatment with Gli-1 siRNA and 5-Fu enhanced the cytotoxic effect on Huh7 cells.
Conclusions:
- Down-regulation of Bcl-2 is crucial for Gli-1 siRNA-induced apoptosis in HCC cells.
- Combination therapy of Gli-1 siRNA and chemotherapy presents a promising strategy for HCC treatment.
- Further in vivo investigations are warranted to explore the clinical potential of these strategies.
