摘要
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
Objective To investigate whether NSAIDs can induce apoptosis of p53 mutant gastric cancer cell line MKN28,to elucidate the regulation of NSAIDs on apoptosis related genes bcl-2 and bax of MKN28.Methods The anti-proliferative effect of NSAIDs was measured by MTT assay.Apoptosis was determined by acridine orange(AO) staining,Annexin-V/PI double staining, laser scanning cytometry(LSC) and flow cytometry (FCM). Alteration of mRNA of bcl-2 and bax genes was detected by reverse transcription polymerase chain reaction (RT-PCR).Results Both indomethacin (Indo) and aspirin (Asp) could inhibit MKN28 gastric cancer cell line growth in a time/dose dependent manner.MKN28 cells appeared slight apoptosis after incubated with Indo 800 μmol/L or Asp 8 mmol/L for 48-96hr, but these tiny differences were not statistically significant. The bax mRNA gradually increased in both Indo and Asp treatment while bcl-2 gene decreased.Conclusion NSAIDs could restrain the proliferation of MKN28,but could not induce notable apoptosis, which indicated mutant p53 gene perhaps blocked NSAIDs induced apoptosis. NSAIDs could promote apoptosis related genes of MKN28 to a pro-apoptosis change,although it couldn't regulate expression of mutant p53 gene.
出处
《肿瘤》
CAS
CSCD
北大核心
2004年第6期570-574,共5页
Tumor
