短发夹RNA沉默血红素氧合酶-1基因表达对胃癌细胞系SGC-7901生物学行为的影响

Effects of heme oxygenase-1 short hairpin RNA transfection on biological behaviors of gastric cancer line SGC7901

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.