研究目的
Investigating the therapeutic effects of targeting survivin expression in cancer cells using a novel 3D survivin promoter-EGFP reporter assay.
研究成果
The novel 3D survivin promoter-EGFP reporter assay demonstrated potential for high-throughput screening of chemicals that down-regulate survivin, a promising molecular target for cancer therapy. The assay showed strong correlation between EGFP expression and survivin transcriptional level, and was effective in evaluating the effects of YM155, doxorubicin, and cisplatin on survivin expression and cell cytotoxicity.
研究不足
The study focused on MCF-7 breast cancer cells and may not be directly applicable to other cancer types. The 3D culture system, while more representative of in vivo conditions, may have limitations in scalability and throughput compared to 2D systems.
1：Experimental Design and Method Selection:
The study developed a novel 3D survivin promoter assay using EGFP as the reporter to assess survivin promoter activity for cancer drug screening. MCF-7 cells were engineered to express EGFP under a human survivin promoter and a CMV promoter.
2：Sample Selection and Data Sources:
Breast cancer MCF-7 cells were used, cultured in 3D polymer-based scaffolds on a 40-microbioreactor platform (40-MBR).
3：List of Experimental Equipment and Materials:
Equipment included a fluorescence plate reader (TECAN GENiosProTM), epifluorescence microscopy system (Nikon Eclipse 80i), and flow cytometry (FACS Calibur). Materials included YM155, doxorubicin, cisplatin, and various cell culture reagents.
4：Experimental Procedures and Operational Workflow:
Cells were cultured in 3D scaffolds, treated with drugs, and EGFP fluorescence was monitored in real-time.
5：Data Analysis Methods:
RT-PCR was used to measure survivin and EGFP mRNA levels, and fluorescence intensity was correlated with cell number and survivin promoter activity.
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