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作 者:陈朝[1] 潘雪珂[1] 余克明[1] Joseph M.Kaminski 庄菁[1]
机构地区:[1]中山大学中山眼科中心眼科学国家重点实验室,510060 [2]美国夏威夷马诺阿生物科学有限公司,96822
出 处:《新医学》2012年第6期376-379,共4页Journal of New Medicine
基 金:国家自然科学基金(30872811)
摘 要:目的:研究川芎嗪对人视网膜母细胞瘤(RB)SO-RB50细胞增殖的影响,并探讨其作用与趋化因子CXCR4表达水平之间的关系。方法:体外培养人SO-RB50细胞,经川芎嗪处理不同时间后,应用MTT法检测SO-RB50细胞的增殖抑制;采用荧光定量逆转录PCR和蛋白免疫印迹法分别从基因和蛋白质水平检测川芎嗪对SO-RB50细胞趋化因子受体CXCR4表达的影响,进而应用流式细胞仪检测SO-RB50细胞的细胞周期分布。结果:MTT结果显示川芎嗪对人SO-RB50细胞增殖有明显的抑制作用,与对照组比较差异具有统计学意义(P<0.01);细胞周期结果显示川芎嗪处理后的SO-RB50细胞G0/G1期增多,S期细胞减少,与对照组比较差异有统计学意义(P<0.01);通过荧光定量逆转录PCR及蛋白免疫印迹实验证明,川芎嗪能下调人SO-RB50细胞CXCR4的表达水平。结论:川芎嗪对人SO-RB50细胞的增殖有明显抑制作用,且能下调SO-RB50细胞中与肿瘤发生及生长密切相关的CXCR4表达水平。Objective: To investigate the effect of tetrame-thylpyrazine(TMP) on cell proliferation,CXCR4 expression,cell cycle phase and apoptosis of human retinoblastoma cell line SO-RB50 and to elucidate its possible molecular mechanisms.Methods: SO-RB50 cells were treated with TMP for 48 hours or 72 hours.Methylthiazolyl tetrazolium assay was performed to test the inhibitory effect of TMP on cell proliferation.Quantitative Real-time(RT-PCR) and western blot were adopted to evaluate CXCR4 expression on SO-RB50 cells.Flow cytometry was used to detect the apoptosis and cell cycle of the SO-RB50 cell line.Results: MTT showed that TMP remarkably inhibited cell viability and growth of SO-RB50 cells(P0.01).RT-PCR and Western blot results showed that TMP down-regulated CXCR4 expression in SO-RB50 cells both at RNA and protein levels.Flow cytometry demonstrated that TMP increased the rate of G0/G1 of SO-RB50 cell cycle(P0.01).Conclusions: TMP can inhibit the viability and growth of human retinoblastoma SO-RB50 cell line.To our knowledge,this is the first study to reveal the mechanism of TMP treatment involving the inhibition of chemokine and CXCR4 expression in SO-RB50.
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