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Objective: use of RNA interference (RNAi) technology design, built for people ER81 gene specific RNAi expression system to study the expression system whether inhibition of human breast cancer cell line MDA-MB-231, ER81 expression to promote breast cancer MDA-MB -231 apoptosis, and provide a new approach for the treatment of breast cancer. Method: 1. ER81 mRNA sequence, screening design, the gene fragment of the synthetic siRNA construct specific siRNA expression vector pGenesill-ETV1A/ETV1B/ETV1C-siRNA random order siRNA and GAPDH-siRNA expression vector as a control, double digested identified and sequenced to verify the correctness of the insertion sequence. 2.MDA-MB-231 cells were divided into transfected recombinant plasmid ETV1A group, transfection recombinant plasmid ETV1B group, recombinant plasmid ETV1C group transfection, blank cell group transfected disorder group (HK) transfected with empty vector group ( KB), GAPDG-siRNA transfected group, liposome method gene stable transfection of human breast cancer cell line MDA-MB-231, semi-quantitative RT-PCR, Western blot assay ER81 mRNA transcription and protein expression levels, and transcription and protein expression observed ER81mRNA suppressed, Flow Cytometry observed siRNA blocking the ER81 gene expression on apoptosis of MDA-MB-231. Results: 1. Successfully constructed for people ER81 mRNA sequence siRNA expression vector and disorder siRNA and GAPDH-siRNA expression vector by restriction analysis and sequencing entirely correct. With liposome recombinant plasmid stably transfected cell line MDA-MB-231, RT-PCR and Western blot the the the detection ETV1A group, ETV1B group ER81 mRNA ETV1C group of breast cancer cells and ER81 protein expression blank cell group, out-of-order group, empty vector group, GAPDH-siRNA group decreased, showed that the ER81 siRNA transfected into cells ER81 gene silencing the which ETV1A Group, ETV1C group gene silencing efficiency than ETV1B group in. 3 compared to the the the flow cytometry ETV1A group, ETV1B group, ETV1C apoptosis of tumor cells and the control group, the difference was not statistically significant. Conclusion: the application expression vector method successfully build ER81 gene-specific siRNA interference expression system pGenesill-ETV1A/ETV1B/ETV1C-siRNA. 2.pGenesill-ETV1A/ETV1B/ETV1C-siRNA liposomes transfected target cells play a gene silencing effect, inhibiting the a ER81mRNA transcription and protein expression to ETV1A, ETV1C gene silencing effect more significantly. 3. The flow cytometry ETV1A group the, ETV1B group ETV1C group rate of tumor cell apoptosis compared with the control group, no statistically significant difference.
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