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Construction of CIAPIN1 -RNAi vector and its effect on drug resistance of breast cancer cells
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Abstract:
Objective : To construct lentiviral expressed vector of siRNA targeting CIAPIN1 and establish breast cancer cells with a stable expression of siRNA- CIAPIN1 , and to investigate the effect of CIAPIN1 on breast cancer cells multi-drug resistance. Methods: The expressed vectors of recombined plasmid of siRNA targeting CIAPIN1 were designed and synthesized. Select the most efficient interfering sequence, spackage, and produce a lentiviral vector with it. Stably transfect CIAPIN1 -RNAi into MCF-7/ADM cells. Detect IC 50 value of different drugs in MCF-7/ADM cells before and after CIAPIN1 interference by MTT. Results: The expressed vectors of recombined plasmid of siRNA targeting CIAPIN1 were successfully synthesized and the most efficient interfering sequence was CIAPIN1 -siRNA1. Stable transfection of CIAPIN1 -RNAi into MCF-7/ADM cells by a lentiviral vector suppressed the expression of CIAPIN1 in MCF-7/ADM cells more than 88%. After RNA interference, IC 50 value of MCF-7/ADM cells to anticancer drugs (paclitaxel, doxorubicin and gemcitabine) significantly decreased from (7.12±0.31), (11.21±1.79), (49.72±4.52) to (1.13±0.06), (4.51±020), (1830±127) μg/ml respectively, suggesting a significant decrease in the drug resisstance of the cells. Conclusion: Lentiviral expressed vector of CIAPIN1 -siRNA can efficiently interfere the expression of CIAPIN1 in MCF-7/ADM cells. The study also confirmed the regulation effect of CIAPIN1 on breast cancer cell multi-drug resistance (MDR).
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Project Supported:
Project supported by Creative Talents in Science and Technology Foundation of Science and Technology Bureau of Harbin （No. 2009RFQXS028）
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