Research On Liver Cancer Cell Drug Resistance、migration And Invasion Potential By Control ABCG2Expression

Objective:In order to further study the expression level of ABCG2cell characteristics and liver cancer stem, design construction of targeted gene cloning vector PcDNA3.1(+)-ABCG2and ABCG2gene silence of ABCG2-siRNA. Research on liver cancer cell drug resistance^migration and invasion potential by control ABCG2expression.Methods:(1)The sequence of CDS gene was amplified by ABCG2, cloned into pcDNA3.1(+) vector.(2)Through the verification method of electrophoresis, spectrophotometer and sequencing was constructed correctly, effect on cell ABCG2to detect the expression of Western blot after transfection.(3) Design and synthesis of three ABCG2-siRNA.(4)Test the inhibitory effect on the expression of ABCG2RT-PCR, Western blot assay after transfection.(5)Tesearch by MTT、Wound Healing and Invasion Assay.Results:1.Construct pcDNA3.1(+)-ABCG2eukaryotic expression plasmid, Through the method of amplification fragment size and fragment size consistent theory, enzyme cut size and size with the theory, positive clone sequencing results are correct, after transfection can significantly increase the expression of ABCG2.2. Design Three siRNA, after transfection inhibited the expression of ABCG2, siRNA2has the highest efficiency.3. Upregulation of ABCG2enhanced the capacity of doxorubicin resistance, migration, and invasion potential.Colclusion:1.Successfully construct pcDNA3.1(+)-ABCG2plasmid;2. Successfully construct ABCG2-siRNA.3. Upregulation of ABCG2enhanced the capacity of doxorubicin resistance, migration, and invasion potential.