摘要
目的研究15-脂氧合酶-1(15-LOX-1)基因在人胃癌细胞AGS中的表达及其对AGS增殖凋亡的影响。方法通过脂质体介导瞬时转染15-LOX-1基因于培养的AGS中,逆转录PCR法和免疫印迹法检测15-LOX-1 mRNA及蛋白表达;四甲基偶氮唑盐法(MTT)、流式细胞术、AnnexinV/PI染色与原位末端标记法(TUNEL)比较转染前后AGS的增殖和凋亡情况。结果胃癌细胞AGS转染后表达有15-LOX-1的mRNA及蛋白。与非转染组及空质粒转染组相比较,转染重组质粒组AGS细胞生长明显受到抑制并且细胞周期停滞在G1期,其凋亡率明显高于非转染组及空质粒转染组。结论15-LOX-1基因能够抑制胃癌细胞AGS的增殖,并促进其凋亡。
Objective To investigate the expression of 15-1ipoxygenase-1 （15-LOX-1） in human gastric cancer cell line AGS and its effects on cell proliferation and apoptosis. Methods 15-LOX-1 gene was transfeeted into AGS cells. The expression of 15-LOX-1 at mRNA and protein levels were measured by reverse transcription polymerase chain reaction（RT-PCR） and western blot. The proliferation, cell cycle and apoptosis of AGS cells were assessed by thiazolyl blue tetrazolium bromide（MTT） assay, flow cytometry,AnnexinV/PI double staining and TdT--mediated dUTP Nick-End Labeling （TUNEL）, respectively. Results RT-PCR and western blot showed that 15-LOX-1 mRNA and protein were only expressed in AGS/15-LOX-1 cells but not expressed in AGS and AGS/peDNA3.1 cells. There was a significant decline in cell proliferation ability of AGS/15-LOX-1 in comparison with control groups revealed by MTT assay. Also, flow cytometry analysis displayed accumulation of cell in the G1 phrase of cell cycle compared with control groups. AnnexinV/PI and TUNEL confirmed that 15-LOX-1 could induce apopto- sis of AGS cells, showing a statistically significant differences in comparison with control groups （P〈 0.01）. Conclusion The expression of 15-LOX-1 can strongly suppress the proliferation and induce apoptosis in human gastric cancer cell line AGS, which provides the experimental evidence for investigating the mechanism and treatment of gastric cancer.
出处
《福建医科大学学报》
2009年第1期23-27,共5页
Journal of Fujian Medical University
基金
福建省自然科学基金(2007J0083)
