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凋亡的作用及与COX 2mRNA、bcl 2、caspase 3凋亡基因蛋白表达的关系 ,以初步探索其诱导凋亡的机制。方法 胃癌细胞的凋亡用电子显微镜、Annexin V FITC染色流式细胞仪技术检测。COX 2mRNA表达用RT PCR法检测。bcl 2和caspase 3蛋白表达用免疫细胞化学技术检测。结果 吲哚美辛在浓度为 5 0 μmol/L时作用 2 4h不能诱导胃癌细胞凋亡 ,作用 4 8、 72h和浓度为 10 0和 2 0 0μmol/L时作用 2 4、 4 8、 72h均可诱导凋亡 ,凋亡率分别为 6 4 8% ,8 2 0 % ;9 14 % ,12 2 7% ,15 11%和 9 95 % ,14 70 % ,19 81% ,呈浓度和时间依赖性。吲哚美辛可降低COX 2mRNA和bcl 2蛋白的表达 ,增加caspase 3蛋白的表达 ,2 0 0 μmol/L吲哚美辛作用 72h ,bcl 2和caspase 3蛋白表达阳性率分别为 (2 2 8± 6 5 ) %和 (31 8± 12 7) % ,对照组为 (44 6± 10 1) %和 (14 8± 6 4 ) % ,与对照组比较 ,差异有极显著性意义 (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表达下调及caspase 3的激活。
Objective To study the mechanisms of indomethacin (NSAID)- in duced apoptosis of human gastric cancer line SGC-7901 by detecting the expressi on of COX-2 mRNA, bcl-2 and caspase-3.Methods Apoptosis of gastric cancer line SGC-7901 was determin ed by electronic microscopy, Annexin V-FITC staining and flow cytometry. The CO X-2 mRNA expression was detected by RT-PCR. The protein expression of bcl-2 a nd caspase-3 was examined by immunohistochemistry. Results Indomethacin of 50 μmol/L at 24 h couldn't induce apop tosis of gastric cancer cells. 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 could induce apoptosis of gastri c cancer cells in a dose- and time-dependent manner. Their apoptotic rates wer e 6.48 %, 8.20 % and 9.14 %, 12.27%,15.11 % and 9.95 %, 14.70 %, 19.81 % respectively. It could increase the caspase-3 expression and decrease the bcl -2 and COX-2 mRNA expression. The positive protein expression rates of bcl-2 and caspase-3 were (22.8±6.5) % and (31.8±12.7) % respectively with indom ethacin of 200 μmol/L at 72 h, while the controls being (44.6±10.1) % and (1 4.8±6.4) % (P<0.01). Conclusion Indomethacin could induce apoptosis in gastric cance r line expressing COX-2 by the downregulation of bcl-2 expression and caspase -3 activation.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2004年第6期720-723,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
