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目的 1.明确非甾体消炎药 (NSAIDs)能否诱导p5 3基因突变的胃癌细胞MKN2 8凋亡。 2 .明确NSAIDs对MKN2 8细胞凋亡相关基因bcl 2、bax表达的调控。方法 1.通过MTT比色法检测NSAIDs对细胞生长活力的影响。 2 .应用丫啶橙染色、Annexin V/PI双染色、共聚焦显微镜、流式细胞术检测细胞凋亡。 3.应用RT PCR(逆转录 聚合酶链反应 )方法检测bcl 2、bax基因水平的改变。结果 1.NSAIDs药物吲哚美辛 (Indo)和阿斯匹林 (Asp)对胃癌细胞株MKN2 8均有生长抑制作用 ,且呈时间 /浓度依赖性增强。 2 .在Indo 80 0 μmol/L、Asp8mmol/L作用 4 8～ 96h后 ,MKN2 8细胞凋亡数量稍有增多 ,但不具有统计学意义。 3.随着药物作用时间的延长 ,MKN2 8细胞的bcl 2基因mRNA表达逐渐减弱 ,bax基因表达逐渐增强。结论 1.NSAIDs可抑制MKN2 8胃癌细胞株增殖。 2 .NSAIDs不能诱导p5 3基因突变的MKN2 8胃癌细胞株发生显著的凋亡 ,提示p5 3基因突变可能阻断了NSAIDs诱导的细胞凋亡。 3.NSAIDs可使MKN2 8细胞凋亡相关基因bcl 2和bax的水平呈现促进凋亡倾向的改变
Objectives 1. To clarify whether non-steroidal anti-inflammatory drugs (NSAIDs) can induce the apoptosis of gastric cancer cell line MKN28 with p5 3 mutation. To clarify the effect of NSAIDs on the expression of bcl 2 and bax in MKN 28 cells. Methods 1. MTT colorimetric assay of NSAIDs on cell viability. Apoptosis was detected by acridine orange staining, Annexin V / PI double staining, confocal microscopy and flow cytometry. 3. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the changes of bcl 2 and bax gene levels. NSAIDs Indo and Asp inhibited the growth of gastric cancer cell line MKN2 8 in a time-and dose-dependent manner. The number of apoptotic MKN2 8 cells increased slightly after treated with Indo 80 0 μmol / L and Asp 8 mmol / L for 48-96 h but not statistically significant. With the prolongation of drug effect, the mRNA expression of bcl 2 in MKN 2 8 cells gradually decreased and the expression of bax gene gradually increased. NSAIDs can inhibit the proliferation of MKN2 8 gastric cancer cell line. NSAIDs could not induce significant apoptosis in MKN2 8 gastric cancer cell line with p5 3 mutation, suggesting that mutation of p5 3 might block NSAIDs-induced apoptosis. NSAIDs can change the levels of apoptosis-related genes bcl 2 and bax in MKN 2 8 cells to promote apoptosis