論文哌立福新阻断Akt对食管癌细胞增殖4.docVIP

哌立福新阻断Akt对食管癌细胞增殖、凋亡影响 罗和生，金灵莉，童强，金曙 （湖北省武汉大学附属人民医院消化内科，武汉市，430000） 摘要 目的：研究不同浓度的哌立福新对食管癌细胞的增殖及凋亡的影响，探讨哌立福新阻断Akt信号通路对食管癌细胞生长的抑制作用。方法：将不同浓度的Akt抑制剂哌立福新加入到人食管癌细胞Eca109中，采用MTT法测定细胞毒性，流式细胞仪测定细胞凋亡率，Western Bloting法检测食管癌细胞中Akt、mTOR、GSK-3β蛋白表达。结果：哌立福新对人食管癌细胞Eca109有明显抑制作用并能诱导食管癌细胞凋亡。MTT结果示，随着浓度的升高、时间延长抑制作用增强，并诱导人食管癌细胞Eca109凋亡，呈剂量-时间效应关系；流式细胞术显示，不同浓度哌立福新作用于人食管癌细胞Eca109 24、48、72、96小时后，凋亡率随着时间和浓度的升高而增大，15μmol/L作用48小时后凋亡率最大为51.3%；Western Bloting示，Akt、mTOR、GSK-3β在人食管癌细胞Eca109中有·表达，哌立福新作用后表达显著降低。结论：哌立福新能显著抑制食管癌细胞增殖并诱导其凋亡。 关键词 哌立福新; 食管癌；凋亡；增殖；Akt;mTOR；GSK-3β Effects of Akt Inhibitor Perifosine on Proliferation and Apoptosis of Esophageal Carcinoma Cells LUO Hesheng, JIN Lingli, TONG Qiang, JIN Shu Department of Digestion,Renmin Hospital,Wuhan University,Wuhan 430000 Abstract Objective:To investigate effects of different concentration of Akt inhibitor perifosine on proliferation and apoptosis of esophageal carcinoma cells。Methods:Eca109 cells were treated with 5、10、15 and 20μmol/L of perifosine,respectively,for 24、48、72、96h。The effect of perifosine on cell proliferation was evaluated by MTT assay。The apoptotic rate of the cells treated with 15μml/L of perifosine was detected by flow cytometry,expressions of Akt、mTOR、GSK-3β were detected with Wstern Bloting。 Results:Perifosine had obvious inhibitory effects on the esophageal carcinoma cells and can significantly induce apoptos in does and time-effective manner。 Apoptotic rate of cells treated with 15μmol/L perifosine for 48h is the most obvious。Expressions of Akt、mTOR、GSK-3 βwere high in esophageal carcinoma cells and they were decreased in cells after treated with perifosine。Conclustions:Perifosine can significantly inhibit the proliferation of esophageal carcinoma cells and induce apoptosis。 Key words Perifosine; Esophageal carcinoma；Apoptosis； Proliferation;Akt;mTOR、GSK-3β 表示。多组间采用随机单位组设计资料的方差分析（two-way ANOVA），P 0.05认为有统计学意义。 2 结果 2.1 MTT测定结果 哌立福新对食管癌Eca109细胞有抑制作用。5