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- W2359014720 abstract "Objective To establish a nasopharyngeal carcinoma(NPC) cell line in which β-catenin expression is stably suppressed,and to investigate the role of Wnt/β-catenin pathway in nasopharyngeal tumorigenesis.Methods Three siRNA interference sequences targeting β-catenin gene CTNNB1 were designed.Double-stranded shRNA hairpins were synthetized and separately cloned into pLKO.1 and pLVTHM vector so as to produce three plasimds of pLKO.1-sh-β-catenin(Experimental group1),pLVTHM-sh-β-catenin(Experimental group2) and pLKO.1-sh-β-catenin-Neg(negative control group).The plasimds and the lentiviral packaging plasmids were co-transfected into the packaging cells 293T,then the supernatant with lentivirus was collected to infect 6-10B.After puromycin selection and culture expansion,stable cell clones were obtained.Intracellular β-catenin and c-myc protein expression was detected by Western blot.The effect of down-regulated expressed β-catenin by RNAi on cell proliferation was quantified by methylthiazoletetrazolium(MTT) assay.Results The total intracellular β-catenin and c-myc proteins were decreased in pLKO.1-sh-β-catenin transfected group.Results from the MTT assay reveaed that the proliferation of β-catenin knockdown cells was significantly decreased(P0.05).The number of cells passing through the membrane of Transwell chamber in the pLKO.1-sh-β-catenin group(23.5±4.6) was lower than the negative control(50.3±5.3,P0.01) and the blank control group(54.3±5.6,P0.01).Conclusion Lentiviral shRNA expression vectors targeting β-catenin gene CTNNB1 were successfully constructed and the vextor could effectively downregulate the expression of β-catenin in 6-10B cells.The Wnt/β-catenin signal transduction pathway was also inhibited.This established NPC cell line may be used as a cell model to further study the β-catenin gene-targeted therapy on nasopharyngeal carcinoma." @default.
- W2359014720 created "2016-06-24" @default.
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- W2359014720 date "2011-01-01" @default.
- W2359014720 modified "2023-09-28" @default.
- W2359014720 title "Construction of a stable human nasopharyngeal carcinoma cell line with down-regulated expression of β-catenin" @default.
- W2359014720 hasPublicationYear "2011" @default.
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