共转染沉默5-LOX/COX-2基因对胃癌细胞SGC-7901生长的影响  

Effect of the 5-LOX/COX-2 gene silencing via cotransfection on the growth of SGC-7901 human gastric cancer cells

作　　者：刘代江[1] 刘纯伦[1] 张瑶[1] 窦娟[1] 

机构地区：[1]重庆医科大学附属第一医院消化内科,重庆400016

出　　处：《解放军医学杂志》2012年第5期444-448,共5页Medical Journal of Chinese People's Liberation Army

基　　金：重庆市卫生局资助课题(05-2-155)~~

摘　　要：目的观察沉默环氧合酶2(COX-2)基因和5-脂氧合酶(5-LOX)基因对胃癌细胞SGC-7901生长的影响。方法分别构建5-LOX/COX-2基因的靶向短发夹RNA(shRNA)真核表达质粒,共同转染胃癌细胞SGC-7901。将细胞分为空白对照组、空质粒组、单独转染组(P-sh5-LOX或P-shCOX-2)及共同转染组(P-sh5-LOX+P-shCOX-2)。分别采用RT-PCR及Western blotting检测COX-2/5-LOX的mRNA及蛋白表达情况,MTT法检测细胞增殖能力,流式细胞术检测细胞凋亡率。结果与空白对照组及空质粒组比较,5-LOX和COX-2基因单独转染组及共同转染组的mRNA和蛋白表达量均受到显著抑制(P<0.05)。P-sh5-LOX组(27.38%±0.64%)、P-shCOX-2组(29.05%±0.61%)及共同转染组(50.23%±1.73%)细胞增殖抑制率均显著高于空质粒组(3.78%±0.39%,P<0.05),共同转染组细胞增殖抑制率显著高于单独转染组(P<0.05)。P-sh5-LOX组、P-shCOX-2组及共同转染组的细胞凋亡率(分别为15.31%±1.53%、16.69%±1.28%、27.27%±1.18%)均显著高于空质粒组(0.99%±0.46%,P<0.05),且共同转染组显著高于单独转染组(P<0.05)。结论同时沉默胃癌细胞SGC-7901的5-LOX及COX-2基因,可显著抑制胃癌细胞增殖,促进其凋亡,其效果优于单独沉默其中一个基因。Objective To observe the effects of cyclooxygenase-2（COX-2） and 5-lipoxygenase（5-LOX） silencing on the growth of SGC-7901 human gastric cancer cells. Methods The eukaryotic expression plasmids of targeted short hairpin RNA containing the 5-LOX and COX-2 genes were constructed for the co-transfection of gastric cancer cells.The cells were divided into control group,plasmid group,alone transfection groups（P-sh5-LOX and P-shCOX-2）,and co-transfection group（P-sh5-LOX＋PshCOX-2）. Reverse transcription polymerase chain reaction and Western blotting analysis were used to detect the COX-2 and 5-LOX mRNA and the protein expression.A 3-（4,5-dimethylazol-2-yl）-2,5-diphenyltetrazolium bromide assay was used to detect cell proliferation,and flow cytometry was used to detect the cell apoptotic rate.Results 5-LOX and COX-2 mRNA and protein expression in alone transfection groups and co-transfection group were significantly lower than those in control group（P0.05）.The rates of SGC-7901 cell proliferation inhibition in P-sh5-LOX group（27.38%±0.64%）,P-shCOX-2 group（29.05%±0.61%）,and co-transfection group（50.23%±1.73%） were significantly higher than that in plasmid group（3.78%±0.39%,P0.05）.The rate of inhibition of cell proliferation in co-transfection group was higher than that in alone transfection groups（P0.05）.The cell apoptosis rates in P-sh5-LOX group（15.31%±1.53%）,P-shCOX-2 group（16.69%±1.28%）,and co-transfection group（27.27%±1.18%） were significantly higher than that in blank plasmid group（0.99%±0.46%,P0.05）.The difference in cell apoptosis rate between co-transfection group and alone transfection groups was statistically significant（P0.05）.Conclusions Simultaneous silencing of the 5-LOX and COX-2 gene in SGC-7901 gastric cancer cells inhibits the proliferation of gastric cancer cells and promotes apoptosis. Cosilencing of 5-LOX and COX-2 produces better results than silencing either gene alone.

关 键 词：基因沉默 胃肿瘤 花生四烯酸盐5-脂氧合酶 前列腺素内过氧化物合酶类 

分 类 号：R735.2[医药卫生—肿瘤]