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作 者:陈洪元[1] 张黎[1] 程力[1] 刘立青[1] 崔彬[2] 苗瑞政[1]
机构地区:[1]山东大学附属省立医院胃肠外科,济南250021 [2]山东大学附属省立医院中心实验室,济南250021
出 处:《中华实验外科杂志》2009年第2期199-201,共3页Chinese Journal of Experimental Surgery
基 金:山东省自然科学基金资助项目(Y2007C127)
摘 要:目的观察血红素氧合酶-1(HO-1)基因沉默对胃癌细胞系SGC-7901生长、增殖的影响。方法构建靶向HO—1的短发夹RNA(shRNA—HO-1)干扰质粒转染人胃癌细胞系SGC-7901,逆转录-聚合酶链反应(RT—PCR)、细胞免疫化学分别在mRNA、蛋白质水平检测抑制效果。流式细胞仪和噻唑蓝(MTT)检测HO-1基因沉默后细胞的细胞周期和生长情况。结果shRNA—HO-1在mRNA、蛋白质水平高效特异地抑制了细胞SGC-7901中HO-1的表达(抑制率分别为62.4%、67.6%);较对照质粒组,HO-1的表达被抑制后,G0/G1期细胞百分比明显减少(52.025±1.638比67.525±1.938,P〈0.05),细胞生长受抑制[2.036±0.072比2.783±0.067(72hA值),P〈0.05]。结论shRNA—HO-1可有效抑制胃癌细胞中HO-1的表达;HO-1的表达减少抑制肿瘤细胞生长。Objective To investigate the effects of heme oxygenase=l short hairpin RNA transfection on biological behaviors of gastric cancer line SGC7901. Methods A eukaryotic expression plasmid of shRNA targeting on HO-1 was constructed and was transiently transfected into human gastric cancer line SGC790L Expression of HO-1mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR)and immunocytochemical stainning. Cell proliferation and cell cycle were determined by MTT and flow cytometry assay. Results HO-1 shRNA effectively inhibited the expression of HO-1 mRNA (62.4%)and protein (67.6%), cell growth [ 2. 036 ±0. 072 vs 2. 783± 0. 067 (72 h), P 〈 0.05 ], and decreased the cells in G0/G1 phase (52. 025± 1. 638 vs 67. 525 ±1. 938, P 〈 0.05 ). Conclusion HO-1 shRNA effectively inhibited the expression of HO-1, thus inhibiting the growth of the tumor cells.
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