Objective : To explore the effect of 5 - Aza - CdR on apoptosis and protein expression alteration in laryngeal squamous carcinoma Hep2. Methods: Hep2 cells were treated with 10 -7 mol/L 5 - Aza - CdR and DMSO respectively. Hoechst33258 staining and flow cytometry were used to detect the effect of 5 - Aza - CdR on Hep2 cell apoptosis. Followed by total protein extraction, two dimensional gel electrophoresis and PDQuest - 7. 0. 1 software together with MALDI - TOF mass spectrometry were used to detect the altered proteins in Hep2 cells. While, western holts were used to confirm the significant difference. Results: Obvious nucleus dense staining were found in Hep2 cells treated with 10 -7mol/L 5 -Aza -CdR for 72 hours, and the apoptosis rate increased from ( 13. 96 ±0. 41 )％ to ( 2. 70 ± 0. 28 )％ , compared with DMSO group , which was significantly different ( P ＜ 0. 05 ). Two dimensional gel electrophoresis detected 5 significantly difference expressed proteins including S100A4. Western bolts further confirmed that 5 - Aza - CdR could significantly up - regulate S100A4 expression in Hep2 cells, which confirmed the two dimensional gel electrophoresis results in our study were credible. Conclusions : Several protein expression alterations mediated by 5 Aza - CdR participate in Hep2 cell apoptosis induced by the same agent directly or indirectly, which providing theoretical fundament for the mechanism of 5 - Aza - CdR exploring treatment in laryngeal squamous carcinoma.%目的:探讨5-Aza-CdR对人喉鳞癌Hep2细胞凋亡及蛋白质表达的影响.方法:Hep2细胞分别经10-7mol/L 5-Aza-CdR和DMSO处理72h,应用Hoechst33258染色及流式细胞仪检测5-Aza-CdR对Hep2细胞凋亡的影响.抽提各组Hep2细胞总蛋白质,行双向电泳,采用PDQuest-7.0.1软件及MALDI-TOF质谱技术筛查5-Aza-CdR对Hep2细胞蛋白表达的影响,并通过Western blot对显著性差异蛋白质进行验证.结果:人喉鳞癌Hep2细胞在10-7mol/L 5-Aza-CdR处理72h后出现明显的细胞核致密浓染,Hep2细胞的凋亡率由对照组的(2.70±0.28)%增加到(13.96±0.41)%,具有统计学意义(P＜0.05).双向电泳筛查了包括S100A4在内的5种表达显著差异的蛋白质,Western blot进一步验证了5-Aza-CdR能明显上调喉鳞癌Hep2细胞中S100A4的表达,进而证明了双向电泳结果的可信性.结论:5-Aza-CdR介导的Hep2细胞部分蛋白质表达改变,直接或间接地参与了5-Aza-CdR诱导的Hep2细胞凋亡的发生,为5-Aza-CdR在喉鳞癌探讨性治疗中的机制研究奠定基础.
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