NSAIDs induced apoptosis of gastric cancer cell lines and the mechanism / 中国癌症杂志

ABSTRACT

Purpose:To investigate whether NSAIDs can induce apoptosis of gastric cancer cell lines,to observe the effect of different p53 genotype on NSAIDs induced apoptosis,to elucidate the regulation of NSAIDs on expression of apoptosis related genes. 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 bcl-2 and bax genes was detected by reverse transcription polymerase chain reaction (RT-PCR).Protein expression was determined by Western-blotting.Results: Indomethacin (Indo) and Aspirin (Asp) inhibited both AGS(wild-type p53)and MKN28(mutant p53) gastric cancer cell lines growth in a time/dose dependent manner.AGS cell line was more sensitive to NSAIDs,which apoptosis percentage was significantly higher than MKN28 under the same condition. The percentages of apoptosis of MKN28 were somewhat higher among NSAIDs treated groups compared with the normal control group,but these slight differences were not statistically significant. The bax mRNA kept increasing since NSAIDs treatment accompanied by a decrease of bcl-2 gene.The Bax protein increased after treatment while the Bcl-2 protein was undetectable, which tendency was more obvious during 6-24hs.Conclusion: Both Indo and Asp could induce apoptosis in gastric cancer cell lines,which adds further theoretical foundation to the anti-cancer use of NSAIDs.NSAIDs could not induce notable apoptosis of MKN28,which indicated the mutant p53 gene perhaps blocked NSAIDs induced apoptosis.One of the major pathways that mediated the anti-tumour response of NSAIDs in gastric cancer cells was through up-regulation of bax and down-regulation of bcl-2 genes and/or proteins.[