摘要
目的 :研究非甾体类抗炎药吲哚美辛诱导人胃癌细胞系SGC 790 1的凋亡作用及对COX 2mRNA表达及c myc、bcl 2、cas pase 3凋亡基因蛋白的表达 ,以探索其凋亡机制。方法 :胃癌细胞的凋亡用电子显微镜、AnnexinV FITC染色流式细胞仪技术测定。COX 2基因表达用RT PCR法测定。c myc、bcl 2和caspase 3蛋白表达用免疫细胞化学技术测定。结果 :吲哚美辛在浓度为 50 μmol/L时作用48、72h和浓度为 1 0 0和 2 0 0 μmol/L时作用 2 4、48、72h均可诱导胃癌细胞凋亡 ,凋亡率分别为 6 .48%、8.2 0 % ;9.1 4 %、1 2 .2 7%、1 5 .1 1 %和 9.95 %、1 4 .70 %、1 9.81 % ,呈浓度和时间依赖性。吲哚美辛可降低COX 2mRNA表达和bcl 2蛋白表达 ,增加c myc和caspase 3蛋白表达。 2 0 0 μmol/L吲哚美辛作用 72h ,bcl 2、c myc和caspase 3蛋白表达阳性率分别为 2 2 .8± 6 .5 %、42 .5± 1 3 .1 %和 31 8± 1 2 .7% ;对照组为 44 .6± 1 0 .1 %、2 4 .74± 9.5 %和 1 4 .8± 6 .4%。两者比较 ,差异有显著性 (P <0 .0 1 )。结论 :吲哚美辛可诱导胃癌细胞SGC 790 1凋亡 ,其机制涉及bcl 2表达下调、c myc表达上调及caspase 3激活。
Purpose: To study the mechanisms of indomethacin - induced apoptosis of human gastric cancer line SGC-7901 by detecting the expressions of COX-2 mRNA, c-myc,bcl-2, caspase-3. Methods: Apoptosis was determined by electronic microscopy, Annexin V-FITC staining and flow cytometry. The mRNA of COX-2 was detected by RT-PCR. The protein expressions of c-myc, bcl-2 and caspase-3 was examined by immunohistochemistry. Results: Indomethacin of 50 μmol/L at 48 and 72 h,and of 100 μmol/L and 200 μmol/L at 24,48 and 72 h induced apoptosis of gastric cancer cells in a dose- and time-dependent manner. Their apoptotic rates were 6.48%, 8.20% and 9.14%, 12.27%, 15.11% and 9.95%, 14.70%, 19.81% respectively. It increased c-myc and caspase-3 expression and decreased bcl-2 and COX-2 mRNA expression. The positive protein expression rates of bcl-2,c-myc and caspase-3 were 22.8±6.5%,42.5±13.1% and 31.8±12.7% respectively with indomethacin of 200 μmol/L at 72 h,while the controls being 44.6±10.1%,24.74±9.5% and 14.8±6.4%(P<0.01). Conclusion: Indomethacin induced apoptosis in gastric cancer line expressing COX-2 by the upregutations of c-myc and the downregulation of bcl-2 expression and caspase-3 activation.This suggested multiple-gene regulations participated in the apoptosis.
出处
《临床消化病杂志》
2003年第6期249-251,253,共4页
Chinese Journal of Clinical Gastroenterology
关键词
胃癌细胞
吲哚美辛
细胞凋亡
凋亡相关基因
Gastric cancer cells Indomethacin Apoptosis Apoptosis-related genes
