过表达trim11对结直肠癌dld-1细胞5-fu敏感性的影响-effect of overexpression of trim 11 on sensitivity of 5 - fu in colorectal cancer dld - 1 cells.docxVIP

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过表达trim11对结直肠癌dld-1细胞5-fu敏感性的影响-effect of overexpression of trim 11 on sensitivity of 5 - fu in colorectal cancer dld - 1 cells.docx

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过表达trim11对结直肠癌dld-1细胞5-fu敏感性的影响-effect of overexpression of trim 11 on sensitivity of 5 - fu in colorectal cancer dld - 1 cells

AbAbstract PAGE PAGE VIABSTRACTObjectives:To preliminarily study the expression of TRIM11 in colorectal cancer tissue or cell and its over-expression in the influence of 5-FU(5-flourouracil) sensitivity. Methods:Collecting the colorectal cancer and its nearby normal fresh postoperation tissue samples and detecting the expression of TRIM11mRNA using the real-time fluorescence quantitative PCR (qRT-PCR).Taking a culture of 11 colorectal cell strains respectively and testing the expression of TRIM11mRNA in each cell strain using the qRT-PCR.Builded the plasmid carrier pSin-TRIM11, and successfully transfected into DLD-1 cell(DLD-1-TRIM11 group) by lentiviruses transduction technique.Empty vector pSin/neo was transfected as control group(DLD-1-vector group). we identified the TRIM11 protein over-expression in DLD-1-TRIM11 group cell by western blot.Then detecting the cell viability in CCK-8 after 24 hours treated with different concentrations of 5-FU respectively.Testing the two group cell apoptosis rate by the flow cytometry after 24 hours treated with 150ug/ml concentration of 5-FU respectively.Forecasting the miRNAs which could regulate TRIM11 through TargetScan Bioinformatics software.Results:Tissue qRT-PCR showed there were five of seven tissue samples which TRIM11mRNA expression level in colorectal cancer tissues were obviously lower than theirs normal tissues(P0.05).Cell qRT-PCR displayed the TRIM11mRNA was different in each cancer cell strain.The expression of HT29 cell was the highest while DLD-1 was the lowest in all 11 colorectal cancer cell strains.TRIM11 protein was over-expression in DLD-1-TRIM11 group than DLD-1-vector group(P0.05).CCK-8 result showed the cell survival activity in the DLD-1-TRIM11 group was quite lower than the control group(P0.05). The half maximal inhibitory concentration(IC50) of DLD-1-TRIM11 group was 13.94ug/ml which was much lower than DLD-1-vector group(IC50=110.24 ug/ml).The flow cytometry result indicated that the cell apoptosis