摘要
目的明确非甾体消炎药(NSAIDs)诱导胃癌细胞凋亡情况和不同p53基因表型对NSAIDs诱导细胞凋亡的影响。方法通过MTT比色法检测NSAIDs对细胞生长活力的影响;应用丫啶橙染色、Annexin-V/PI双染色、共聚焦显微镜、TUNEL法检测细胞凋亡;应用流式细胞术进行凋亡细胞计数。结果NSAIDs药物吲哚美辛(Indo)和阿斯匹林(Asp)对胃癌细胞株AGS(p53+/+)、MKN28(p53-/-)均有显著的生长抑制作用,且呈时间依赖性增强。一定浓度的NSAIDs作用一定时间后,AGS、MKN28细胞均发生凋亡的形态学和生化学改变。在相同作用条件下,AGS细胞凋亡率明显高于MKN28细胞。处理组MKN28细胞凋亡数量虽有所增多,但不具有统计学意义 结论NSAIDs可诱导胃癌细胞凋亡,AGS细胞对NSAIDs更为敏感,Indo的凋亡诱导作用更为强烈。p53基因突变可能阻断了NSAIDs绣导的细胞凋亡。
Objective To investigate whether NSAIDs can induce, apoposis of gastric cancer cell lines, and to observe the effect of different p53 genotype on NSAIDs induced apoptosis. Methods The anti-proliferative effect of NSAIDs was measured by MTT assay. Apoptosis was observed by acridine orange(AO) staining, Annexin-V/PI double staining, laser scanning confocul microscopy (LSC) and TdT-mediated dUTP nick end labelling assay (TUNEL). Flow cytometry (FCM) was used to count apoptotic cells. Results Indomethacin (Indo) and aspirin (Asp) inhibited both AGS( wild-type p53) and MKN28(mutant p53) gastric cancer cell lines growth in a time-dependent manner. Apoptosis appeared both AGS and MKN28 cells after incubation with Indo 800 μM/Asp 8 mmoL/L for 24 h or Indo 400μmoL/L for 48 h, while FCM result indicated that AGS cell line was more sensitive to NSAIDs, with the apoptosis percentage significantly higher than MKN28. The apoptosis percentage of MKN28 was somewhat higher among NSAIDs treated groups compared with the control group, but these slight differences were not statistically significant. Conclusion Both Indo and Asp could induce apoptosis in gastric cancer cell lines. Apoptosis was more notable in Indo treated AGS cell line. The mutant p53 gene perhaps blocked NSAIDs induced apoptosis.
出处
《上海第二医科大学学报》
CSCD
2004年第6期432-435,共4页
Acta Universitatis Medicinalis Secondae Shanghai
