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机构地区:[1]三峡大学第一临床医学院肝胆脾胰外科,湖北宜昌443002
出 处:《基础医学与临床》2014年第7期974-978,共5页Basic and Clinical Medicine
摘 要:目的探讨小分子干扰RNA(siRNA)沉默衰亡加速因子(CD55)基因对胰腺癌细胞增殖的影响。方法针对CD55基因设计3对siRNA,转染胰腺癌BxPC-3细胞,用实时定量PCR和Western blot检测CD55 mRNA和蛋白的表达,并用流式细胞术检测细胞生物学行为的改变。结果转染BxPC-3细胞48 h后,3条siRNA中仅siRNA3能显著抑制CD55 mRNA和蛋白的表达,细胞凋亡比率显著增加(P<0.05);细胞增殖率显著减少(P<0.05);细胞G1/G0期升高,G2期变化不明显,S期下降(P<0.05)。结论使用siRNA能够有效地沉默CD55基因的表达,并显著影响胰腺癌细胞的生物学行为。Objective To study the specific effect of silencing of CD55 gene by small interfering RNA (siRNA) on the growth of human pancreatic cancer cell line BxPC-3.Methods Three CD55-specific siRNAs and one negative siRNA were designed and then transfected into human pancreatic cancer cell line BxPC-3 by Lipofectamine 2000.Quantitative real-time PCR(qRT-PCR) and Western blot were used to check the expression of CD55 mRNA and protein respectively.And flow cytometry were used to detect apoptosis,proliferation and cell cycle of target cells.Results One of three CD55-specific siRNAs,siRNA3 silenced the expression of the CD55 gene.The expression of CD55 mRNA and protein remarkably decreased.The cells transfected with siRNA3 were found to increase apoptosis and reduce proliferation significantly (P 〈 0.05) ; the target cells stagnated in phase G1 after transfection (P 〈 0.05).Conclusions siRNA-mediated silencing of CD55 can effectively affect the biological behavior of human pancreatic cancer cell line BxPC-3.
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