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Effects of siRNA-Mediated Knockdown of HDAC1 on the Biological Behavior of Esophageal Carcinoma Cell Lines

机译:siRNA介导的HDAC1基因敲低对食管癌细胞系生物学行为的影响

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BACKGROUND HDAC1 has been shown to be closely associated with the occurrence of tumors. We aimed to investigate the effects of siRNA-mediated HDAC1 knockdown on the biological behavior of esophageal carcinoma cell lines. MATERIAL AND METHODS HDAC1 expression in esophageal cancer cell lines TE-1, Eca109, and EC9706 was compared by Western blot analysis. These cells were transfected with siRNA-HDAC1 and cell proliferation was evaluated by MTT assay to select the optimum cell line for subsequent experiments. The effects of siRNA-HDAC1 on the migration and invasion of the selected cell line were assessed by transwell assay. The expression of cell cycle-related proteins cyclinD1, p21 and p27, and epithelial-mesenchymal transition (EMT)-related protein zonula occludens-1 (ZO-1), E-cadherin and vimentin was determined by Western blot analysis. RESULTS HDAC1 expression in TE-1, Eca109 and EC9706 cells was significantly higher compared with normal esophageal cell line HEEC (P<0.01). MTT assay, Western blot and RT-PCR analyses demonstrated that the inhibitory effects of siRNA on HDAC1 expression and cell viability in TE-1 cells were the highest among all cell lines, which was therefore used in subsequent experiments. After TE-1 cells were transfected with siRNA-HDAC1, their migration and invasion were significantly lower compared with the controls (P<0.01). CyclinD1 and vimentin expression was significantly lower compared with the controls (P<0.01), whereas the expression of p21, p27, ZO-1 and E-cadherin was significantly higher (P<0.01). CONCLUSIONS The siRNA-mediated HDAC1 knockdown significantly inhibited the proliferation, migration and invasion of TE-1 cells probably by regulating the expression of cell cycle- and EMT-related proteins.
机译:背景技术已经证明HDAC1与肿瘤的发生密切相关。我们旨在研究siRNA介导的HDAC1敲低对食管癌细胞系生物学行为的影响。材料与方法通过蛋白质印迹分析比较了食管癌细胞系TE-1,Eca109和EC9706中HDAC1的表达。将这些细胞用siRNA-HDAC1转染,并通过MTT分析评估细胞增殖,以选择最佳细胞系进行后续实验。通过transwell分析评估了siRNA-HDAC1对所选细胞系迁移和侵袭的影响。通过蛋白质印迹分析确定细胞周期相关蛋白cyclinD1,p21和p27以及上皮-间质转化(EMT)相关蛋白小带闭塞蛋白-1(ZO-1),E-钙黏着蛋白和波形蛋白的表达。结果与正常食管细胞HEEC相比,TE-1,Eca109和EC9706细胞中HDAC1的表达显着升高(P <0.01)。 MTT分析,Western印迹和RT-PCR分析表明,siRNA对TE-1细胞中HDAC1表达和细胞活力的抑制作用在所有细胞系中最高,因此可用于后续实验。 siRNA-HDAC1转染TE-1细胞后,其迁移和侵袭能力明显低于对照组(P <0.01)。与对照组相比,CyclinD1和波形蛋白的表达显着降低(P <0.01),而p21,p27,ZO-1和E-cadherin的表达显着升高(P <0.01)。结论siRNA介导的HDAC1敲低可能通过调节细胞周期和EMT相关蛋白的表达来显着抑制TE-1细胞的增殖,迁移和侵袭。

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