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DNA Dual-Color Probes for Real-Time Monitoring of p21 mRNA and Bax mRNA during Apoptosis in Breast Cancer Cells
Xiao Qiao Wu1, Chun Tian1, Yiaobo Zhang1
1Key Laboratory of Luminescence Analysis and Molecular Sensing (Southwest University), Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, P. R. China.
Abstract:
Monitoring apoptosis in breast cancer cells is crucial for understanding the molecular mechanisms underlying breast cancer and evaluating the therapeutic effects of anticancer drugs. Real-time monitoring of p21 mRNA and Bax mRNA during apoptosis offers a valuable tool for assessing drug efficacy. Tetrahedral DNA frameworks (TDFs) are known for their ease of synthesis and multiple signal outputs. Leveraging these properties, this study developed DNA dual-color probes (DDCPs) for the real-time monitoring of apoptosis in breast cancer cells. In the presence of p21 and Bax mRNA, strand displacement reactions within the DDCPs restore Cy5 and FAM fluorescence signals, respectively. This strategy is consistent with the results of RT-qPCR studies. Moreover, the DDCPs allow for the specific investigation of apoptotic pathways in breast cancer cells influenced by different drugs by using distinct fluorescent signals for pathway differentiation. This method holds promise as a powerful tool for investigating apoptotic mechanisms in breast cancer cells, uncovering associated signaling pathways, and screening potential therapeutic drugs.
Insights
This study introduces DNA dual-color probes (DDCPs) for real-time apoptosis monitoring in breast cancer cells. These probes track p21 and Bax mRNA, aiding in evaluating anticancer drug efficacy and understanding apoptosis.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Monitoring apoptosis in breast cancer is vital for understanding disease mechanisms and drug responses.
- p21 and Bax mRNA levels are key indicators during apoptosis, crucial for assessing therapeutic effects.
Purpose of the Study:
- To develop and validate DNA dual-color probes (DDCPs) for real-time monitoring of apoptosis in breast cancer cells.
- To differentiate and investigate specific apoptotic pathways affected by various anticancer drugs.
Main Methods:
- Utilized tetrahedral DNA frameworks (TDFs) to create DDCPs capable of multiplexed fluorescence signaling.
- Employed strand displacement reactions within DDCPs to detect p21 and Bax mRNA, restoring Cy5 and FAM fluorescence.
- Validated DDCP results against established RT-qPCR methods.
Main Results:
- DDCPs successfully enabled real-time monitoring of p21 and Bax mRNA during apoptosis in breast cancer cells.
- The probes demonstrated distinct fluorescence outputs for specific mRNA targets, allowing pathway differentiation.
- Results correlated well with RT-qPCR, confirming probe accuracy and reliability.
Conclusions:
- DDCPs offer a sensitive and specific method for real-time apoptosis monitoring in breast cancer.
- This approach facilitates the investigation of drug-induced apoptotic pathways and aids in therapeutic drug screening.
- The technology shows significant promise for advancing breast cancer research and drug development.
