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Survivin--an attractive target for RNAi in non-Hodgkin's lymphoma, Daudi cell line as a model
Gu Congmin1, Zeng Mu, Ma Yihui
1Department of Pathology, Guangzhou Women and Children Hospital, Guangzhou, PR China.
Abstract:
RNA interference (RNAi) has been widely used in tumor gene therapy, antivirus and gene drug selection. Survivin gene is highly expressed in non-Hodgkin's lymphoma (NHL) tissues and high malignancy Burkitt's lymphoma cell line-Daudi and it is regarded as a potential target of gene therapy for NHL. This study used a vector-based short hairpin RNA (shRNA) technique to explore the effect of RNAi-mediated survivin gene silencing on apoptosis and proliferation of Daudi cells. Recombinant plasmid survivin-shRNA was transfected into Daudi cells transiently and stably. The expression of survivin was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The apoptosis of Daudi cells after transfection were evaluated by flow cytometry. After transfection of survivin-shRNA, the levels of survivin mRNA were significantly reduced by 64.20% (transient transfection) and 62.32% (stable transfection), respectively; The levels of survivin protein were significantly reduced by 63.50% (transient transfection) and 61.88% (stable transfection); compared with control-shRNA and PBS treated groups. Apoptosis of Daudi cells were significantly higher in the transfection group than in the control group, respectively 21.30 +/- 2.96% (transient transfection) and 19.10 +/- 2.15% (stable transfection). In conclusion, it was suggested that survivin could be an attractive target for new anti-cancer intervention of NHL and vector-based survivin-shRNA could effectively reduce the expression of survivin and induce cell apoptosis and growth inhibition of NHL cells.
Insights
RNA interference targeting the survivin gene effectively reduces its expression in non-Hodgkin's lymphoma cells. This gene silencing induces apoptosis and inhibits proliferation, suggesting survivin as a therapeutic target for NHL.
Area of Science:
- Molecular Biology
- Cancer Research
- Gene Therapy
Background:
- Survivin gene is highly expressed in non-Hodgkin's lymphoma (NHL) tissues, particularly in high-grade Burkitt's lymphoma cell line-Daudi.
- Survivin is recognized as a potential therapeutic target for NHL due to its role in cancer progression.
Purpose of the Study:
- To investigate the efficacy of RNA interference (RNAi)-mediated survivin gene silencing on apoptosis and proliferation of Daudi cells.
- To evaluate the use of vector-based short hairpin RNA (shRNA) for survivin gene silencing in NHL cells.
Main Methods:
- Transient and stable transfection of Daudi cells with recombinant plasmid survivin-shRNA.
- Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot to assess survivin expression levels.
- Flow cytometry to evaluate the apoptosis rate in Daudi cells post-transfection.
Main Results:
- Survivin mRNA levels were significantly reduced by approximately 64% (transient) and 62% (stable transfection).
- Survivin protein levels decreased by approximately 63.5% (transient) and 61.9% (stable transfection) compared to controls.
- Apoptosis rates increased significantly in survivin-shRNA transfected cells (21.30% transient, 19.10% stable) compared to control groups.
Conclusions:
- Survivin is a promising therapeutic target for novel anti-cancer interventions in NHL.
- Vector-based survivin-shRNA effectively suppresses survivin expression, inducing apoptosis and inhibiting growth in NHL cells.
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