地西他滨诱导人食管癌细胞EC8712凋亡的作用机制研究  

MECHANISMS OF DICITABINE-INDUCED APOPTOSIS IN HUMAN ESOPHAGEAL CANCER CELL LINE EC8712

作　　者：黄丽[1] 范宇洋[1] 张天若[2] Huang Li;Fan Yuyang;Zhang Tiatiruo(Department of General Oncology,Second Affiliated Hospital of Shenyang Medical College,Shenyang 100031,China)

机构地区：[1]沈阳医学院附属第二医院普外肿瘤科,辽宁省沈阳市100031 [2]中国医科大学附属第四医院核医学科

出　　处：《中国煤炭工业医学杂志》2020年第4期352-356,共5页Chinese Journal of Coal Industry Medicine

基　　金：辽宁省自然科学基金(编号:20180550506)。

摘　　要：目的探究地西他滨(DAC)诱导人食管癌细胞EC8712凋亡的作用及相应机制。方法不同浓度DAC作用于人食管癌细胞EC8712,MTT法检测DAC对EC8712细胞的抑制作用;流式细胞术检测DAC对EC8712细胞凋亡及周期的影响;DNA凝胶电泳检测DNA变化情况;Western blot法检测Bax、Bcl-2蛋白表达。结果DAC对EC8712细胞抑制作用随着孵育时间延长及DAC浓度升高而增强,具有时间和浓度依赖性。DAC处理人食管癌细胞EC8712 DNA后细胞凋亡率显著高于对照组(P<0.05),S期及G2期细胞占比显著高于对照组(P<0.05)。DNA凝胶电泳结果显示对照组细胞不出现DNA降解现象,经DAC处理后细胞形成180~200bp的整数倍的寡核苷酸片段,在凝胶电泳上表现为梯形电泳图谱(DNA ladder)。DAC处理后EC8712细胞Bax蛋白表达及Bax/Bcl-2比值高于对照组(P<0.05),Bcl-2蛋白表达低于对照组(P<0.05)。结论DAC对人食管癌细胞EC8712具有增殖抑制作用,通过将细胞阻滞于S期及G2期,上调Bax蛋白表达,下调Bcl-2蛋白表达,诱导人食管癌细胞EC8712凋亡。Objective To investigate the apoptotic effect and mechanism of dexitabine(DAC)on human esophageal cancer cell line EC8712.Methods Different concentrations of DAC acted on human esophageal cancer cells EC8712,MTT method was used to detect the inhibitory effect of DAC on EC8712 cells;flow cytometry was used to detect the effect of DAC on EC8712 cell apoptosis and cycle;DNA gel electrophoresis was used to detect DNA changes;Western blot method was used to detect the expression of Western and protein.Results The inhibitory effect of DAC on EC8712 cells increased with the prolongation of incubation time and the increase of DAC concentration in a time-dependent and concentration-dependent manner.The apoptotic rate of EC8712 DNA treated with DAC was significantly higher than that of control group(P<0.05),and the proportion of S and G2 cells was significantly higher than that of control group(P<0.05).DNA gel electrophoresis showed that there was no DNA degradation in the control group.After DAC treatment,the cells formed an integer multiple oligonucleotide fragment of 180~200 bp,and showed ladder electrophoresis pattern on the gel electrophoresis(DNA ladder).The expression of Bax protein and Bax/Bcl-2 ratio in EC8712 cells treated with DAC were higher than those in control group(P<0.05),and the expression of Bcl-2 protein was lower than that in control group(P<0.05).Conclusion DAC can inhibit the proliferation of human esophageal cancer cell EC8712.By blocking cells at S and G2 stages,up-regulation of Bax protein expression and down-regulation of Bcl-2 protein expression can induce apoptosis of human esophageal cancer cell EC8712.

关 键 词：地西他滨 人食管癌细胞 EC8712 凋亡 

分 类 号：R735.1[医药卫生—肿瘤]