索拉菲尼调控Y盒结合蛋白1抑制肾细胞癌增殖、趋化和侵袭的初步探讨
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国家自然科学基金青年基金(81202380和81402094);天津市应用基础与前沿技术研究计划青年基金项目(14JCQNJC11600);中国博士后基金面上项目(2013 M541189)。


Preliminary study of inhibitory effects of sorafenib for proliferation, chemotaxis and invasion of renal cell carcinoma by regulating Y-box binding protein
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    摘要:

    摘要:目的:探讨索拉菲尼在调控Y盒结合蛋白1(YB1)表达以及影响肾癌细胞系SW839的增殖、趋化和侵袭中的作用。 方法:构建慢病毒介导的YB1干扰载体pLKO.1-shYB1,建立YB1降表达的稳定SW339细胞系;用不同浓度的索拉菲尼处理SW839细胞后,western blot验证YB1的干扰效率以及索拉菲尼对YB1蛋白质表达水平的影响;MTT实验、Transwell趋化实验和Matrigel侵袭实验分别检测YB1基因敲减和索拉菲尼处理对SW839细胞增殖、趋化和侵袭能力的影响。 结果:成功构建YB1降表达的稳定细胞系SW839-shYB1;western blot检测结果发现,SW839-shYB1组YB1蛋白表达水平低于SW839-scr(阴性对照)组和SW839组(P<0.05);与SW839组相比,4、8 μmol/L索拉菲尼处理组YB1蛋白质的表达水平均下降(P<0.05)。MTT实验结果表明,敲减YB1基因(t分别为6.13、4.61和17.18)或经8、12、16 μmol/L索拉菲尼作用后(t分别为4.04、7.54和16.38)SW839细胞的增殖能力降低(P均<0.05);Transwell结果表明,与对照组比较,经表皮生长因子(EGF)诱导后,SW839 shYB1组和索拉菲尼处理组的细胞趋化能力降低(t分别为11.52和17.94,P均<0.05),侵袭能力亦降低(t分别为6.64和4.83,P均<0.05)。 结论:YB1在调节肾癌细胞系SW839的增殖、趋化和侵袭中起重要作用,索拉菲尼可能通过抑制SW839细胞中YB1表达来抑制肾癌细胞的增殖、趋化和侵袭。

    Abstract:

    Abstract: Objective:To investigate the effect of sorafenib on proliferation, chemotaxis and invasion of renal carcinoma cell SW839 through modulating the expression of Y-box binding protein 1(YB1). Methods:Lentivirus mediated YB1 shRNA knockdown vector (pLKO.1-shYB1) was constructed and renal carcinoma cell line with stable YB1 knockdown (SW839-shYB1) was established. SW839 cells were treated with sorafenib in various concentrations. The knockdown efficiency of YB1 and the effect of sorafenib on YB1 expression were confirmed by western blot. The MTT, Transwell chemotaxis and Matrigel invasion assay were used to evaluate the role of YB1 and sorafenib on SW839 cell proliferation, chemotaxis and invasion respectively. Results:Renal cell line SW839-shYB1 with stable YB1 knockdown was successfully constructed. Western blot showed significant decrease of YB1 protein in SW839 shYB1 group compared with SW839 scr and SW839 group (P<0.05). YB1 expression was significantly down regulated in SW839 treated with 4 or 8 μmol/L sorafenib compared with SW839 control group (P<0.05). MTT assay showed the proliferation rate of SW839 was reduced (P<0.05) after the 1, 2 and 3 day of YB1  knockdown (t=6.13, 4.61 and 17.18, respectively) or 8, 12 and 16 μmol/L sorafenib treatment (t=4.04, 7.54 and 16.38, respectively). Transwell results showed that epidermal growth factor (EGF)-induced cell chemotaxis ability in SW839-shYB1 group and sorafenib treated group were decreased compared with control group (t=11.52 and 17.94, P<0.05), invasion ability was also reduced (t=6.64 and 4.83, P<0.05). Conclusion:YB1 may play an important role in modulating SW839 cell proliferation, chemotaxis and invasion. Sorafenib should be able to inhibit the expression of YB1 so as to suppress SW839 cell proliferation, chemotaxis and invasion.

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陈雅婧,付东鹤,王勇,刘运德,岳丹.索拉菲尼调控Y盒结合蛋白1抑制肾细胞癌增殖、趋化和侵袭的初步探讨[J].临床检验杂志,2015,(11):849-853

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  • 收稿日期:2015-10-19
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  • 在线发布日期: 2016-02-29
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