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Effect of RNA interference of iASPP on the apoptosis in MCF-7 breast cancer cells
1Laboratory of International Cooperation, Southwest Hospital, Third Military Medical University, Chongqing, PRC. zejunliu2000@yahoo.com.cn
Abstract:
ASPP family is proved to be apoptotic specific regulators of p53. Among them, iASPP acts as an inhibitor of p53. To investigate the effect of the iASPP RNAi on the apoptosis of breast cancer cell MCF-7, we transfected the recombinant plasmid PGCsilencer H1/Neo/GFP/RNAi into MCF-7. The iASPP expression was analyzed by RT-PCR and Western blot. The cell apoptosis was detected by FCM. The results show that the expression of iASPP is descended and the apoptosis rate is increased after transfection. Therefore, we conclude that inhibition of expression of iASPP may resume the ability of p53 to induce apoptosis in MCF-7 cells.
Insights
Inhibiting iASPP expression in breast cancer cells (MCF-7) increases apoptosis. This suggests iASPP RNAi may restore the tumor suppressor p53
Area of Science:
- Molecular Biology
- Cancer Research
- Cell Biology
Background:
- The ASPP (Apoptosis-Stimulating Protein of p53) family regulates p53-mediated apoptosis.
- iASPP (inhibitory ASPP) functions as a negative regulator of p53.
- Understanding iASPP's role is crucial for breast cancer therapy.
Purpose of the Study:
- To investigate the effect of iASPP RNA interference (RNAi) on apoptosis in MCF-7 breast cancer cells.
- To determine if inhibiting iASPP can re-sensitize cancer cells to apoptosis.
Main Methods:
- MCF-7 cells were transfected with a recombinant plasmid for iASPP RNAi.
- iASPP expression levels were quantified using RT-PCR and Western blotting.
- Apoptosis rates were assessed via Flow Cytometry (FCM).
Main Results:
- Successful transfection led to a significant decrease in iASPP expression.
- A notable increase in the apoptosis rate of MCF-7 cells was observed post-transfection.
- These findings correlate iASPP inhibition with enhanced cancer cell death.
Conclusions:
- Inhibition of iASPP expression via RNAi can induce apoptosis in MCF-7 breast cancer cells.
- Restoring p53's apoptotic function by targeting iASPP is a potential therapeutic strategy.
- Targeting iASPP offers a promising avenue for breast cancer treatment development.