摘要
目的研究15-LOX-1基因对人胃癌细胞增殖的影响。方法通过脂质体介导瞬时转染15-LOX-1基因于培养的胃癌细胞株(AGS)中,以转染空载体pcDNA3.1(+)的细胞及未转染质粒的细胞作为对照;用RT-PCR和Western blot检测人胃癌细胞的15-LOX-1 mRNA及蛋白表达;倒置显微镜观察转染前后AGS形态变化;用MTT法比较转染前后AGS的增殖水平,流式细胞术检测转染后AGS细胞周期的变化。结果胃癌细胞系中未检测到15-LOX-1mRNA和蛋白的表达,转染后的AGS表达有15-LOX-1的mRNA及蛋白。转染重组质粒组细胞增殖显著低于未转染组及空质粒转染组(P<0.05),转染后AGS细胞G0/G1期细胞明显增加,S期细胞明显减少,与未转染组及空质粒转染组相比均有显著性差异(P<0.05)。结论15-LOX-1基因能抑制胃癌细胞的增殖,为进一步探讨胃癌的发病机制及治疗提供实验依据。
Objective To investigate the effects of 15-LOX-1 on the proliferation of human gastric caner AGS cells. Methods AGS cells were divided into three groups： AGS/15-LOX-1 group transfected with pcDNA3.1 （ ＋ ）/15-LOX- 1 , vector control group transfected with peDNA3.1 （ ＋ ） and blank control group without transfeetion. Plasmids of pcDNA3.1 （ ＋ ）/15-LOX-1 encoding 15-LOX-1 gene was transiently transfected into AGS cells with Transfast Reagent. RT- PCR and Western blot were used to detect the expression of 15-LOX-1 mRNA and protein respectively. Then light microscopy was used to measure the effects of 15-LOX-1 expression on cell morphology. After transfeetion, MTT assay was applied to evaluate cell proliferation viability and flow eytometry was used to detect the cell cycle distribution of AGS cells. Results RT-PCR and Western blot showed that 15-LOX-1 mRNA and protein were expressed in AGS/15-LOX-1 cells compared with control groups. The MTT results indicated that AGS/15-LOX-1 group exhibited a slower growth rate compared with control groups（ P 〈0.05 ）. Also, flow cytometry analysis revealed that in AGS/15-LOX-1 ,the number of cells in G0/G1 phrase increased markedly, while the S phrase decreased compared with control groups （P 〈 0.05）. Conclusion The expression of 15-LOX-1 can suppress the proliferation of human gastric cancer AGS cells, which provides the experimental evidence for investigating the mechanism and treatment of gastric cancer.
出处
《胃肠病学和肝病学杂志》
CAS
2008年第9期753-757,共5页
Chinese Journal of Gastroenterology and Hepatology
基金
福建省自然科学基金资助项目(2007J0083)
