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Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death
Published on: October 21, 2012
Gene expression pattern in apoptotic QGY-7703 cells induced by homoharringtonine
Wei Jin1, Le-Feng Qu, Qin Chen
1Breast Cancer Institute, Cancer Hospital, Department of Oncology, Shanghai Medical College, Institute of Biomedical Science, Fudan University, Shanghai 200032, China. zhimingshao@yahoo.com
Aim:
To classify the genes responsible for apoptosis in QGY-7703 cells induced by homoharringtonine (HHT).
Methods:
Apoptosis in QGY-7703 cells induced by HHT was demonstrated by DNA fragmentation and morphological observation. cDNA microarray technology was used to detect gene transcription, and the result of microarrays for genes was confirmed by RT-PCR.
Results:
Seventy-eight individual mRNA were identified and their transcription levels changed significantly. Those genes, of which 68% were upregulated and 32% were downregulated, were partially related to apoptosis. They were mostly oncogenes, tumor suppressors, enzymes, and kinases.
Conclusion:
HHT is a potential drug in the treatment of liver cancer. TGF-beta, TNF, FAS, p38MAPK, and p53 apoptosis signaling pathways were activated during apoptosis in QGY-7703 cells. Such inducible genes may play important roles in apoptosis and deserve to be further studied.
Insights
Homoharringtonine (HHT) induces apoptosis in liver cancer cells by altering gene expression. Key apoptosis signaling pathways were activated, indicating HHT
Area of Science:
- Molecular Biology
- Cell Biology
- Cancer Research
Background:
- Liver cancer remains a significant global health challenge.
- Identifying novel therapeutic agents and understanding their mechanisms of action is crucial.
- Homoharringtonine (HHT) has shown promise as an anti-cancer agent.
Purpose of the Study:
- To identify genes involved in apoptosis induced by HHT in QGY-7703 liver cancer cells.
- To elucidate the molecular mechanisms underlying HHT-induced cell death.
Main Methods:
- Induction of apoptosis in QGY-7703 cells using HHT.
- Assessment of apoptosis via DNA fragmentation and morphological changes.
- Global gene expression profiling using cDNA microarray technology.
- Validation of gene expression changes using Reverse Transcription Polymerase Chain Reaction (RT-PCR).
Main Results:
- Significant alterations in the transcription levels of 78 individual mRNAs were observed.
- 68% of affected genes were upregulated, and 32% were downregulated.
- Affected genes included oncogenes, tumor suppressors, enzymes, and kinases, many related to apoptosis.
Conclusions:
- HHT demonstrates potential as a therapeutic drug for liver cancer treatment.
- Activation of TGF-beta, TNF, FAS, p38MAPK, and p53 signaling pathways was confirmed.
- Inducible genes identified in this study warrant further investigation for their role in HHT-induced apoptosis.
