摘要
目的:探讨PIK3CA在食管鳞癌细胞系Eca109中的作用.方法:用脂质体介导siRNA瞬时转染Ecal09细胞,转染细胞分为3组:空白对照组(正常培养的Eca109细胞)、阴性对照组(转染随机序列的siRNA)及实验组(转染PIK3CA-siRNA).运用Western blot技术检测PIK3CA基因干扰后蛋白水平表达的变化;四甲基偶氮唑盐微量酶反应比色法(MTT法)和细胞划痕实验检测PIK3CA干扰后Eca109细胞增殖和迁移能力的改变;流式细胞仪检测PIK3CA干扰后细胞周期和凋亡率的变化.结果:干扰PIK3CA基因表达后,其蛋白水平表达较空白对照组及阴性对照组显著性降低(P<0.05).MTT实验和划痕实验显示干扰PIK3CA的表达后,Eca109细胞增殖受到显著性抑制(P<0.05),迁移能力显著性降低(P<0.05).流式细胞仪检测下调P I K3C A的表达后细胞周期阻滞在细胞分裂的S期(P<0.05),且使细胞凋亡率显著性增加(P<0.05).结论:PIK3CA基因能够促进Eca109细胞的增殖和迁移能力,并具有抗凋亡作用.PIK3CA基因有望成为食管鳞癌治疗的潜在靶点.
AIM： To explore the role of PIK3CA in cell pro-liferation, migration and apoptosis in esopha- geal squamous cell carcinoma （ESCC） cell line Ecal09.METHODS： Ecal09 cells were divided into three groups： a blank control group （non-transfected cells）, a negative control group （transfected with a scramble siRNA） and an experimental group （transfected with a specific siRNA against PIK3CA）. After transfection using Lipofectamin 2000, the protein expression of PIK3CA was detected by Western blot. Cell proliferation and migration were examined by MTT assay and wound healing assay, respectively. Cell apopto- sis and cell cycle progression were analyzed by flow cytometry. RESULTS： After transfection with the specific siR- NA against P1K3CA, PIK3CA protein expression was significantly decreased compared with the blank control group and negative control group （P 〈 0.05）. After successful knock-down of PIK3CA, both cell proliferation and migration were signifi- cantly reduced （P 〈 0.05）, the cell cycle was ar- rested at S phase （P 〈 0.05）, and the apoptosis rate was significantly increased （P 〈 0.05）. CONCLUSION： PIK3CA plays an important role in promoting cell proliferation and migration and suppressing apoptosis, which may become a new potential target in the therapy of metastasis of ESCC.
出处
《世界华人消化杂志》
北大核心
2014年第32期4893-4900,共8页
World Chinese Journal of Digestology
基金
新疆维吾尔自治区科技援疆基金资助项目
No.201191157
国家自然科学基金资助项目
Nos.81160303
81260359
U1303321
新疆乌鲁木齐感染与肿瘤重点实验室(培育基地)开放课题基金资助项目
No.WIT-2013-03~~
关键词
食管鳞癌
PIK3CA
SIRNA
增殖
迁移
细
胞凋亡
Esophageal squamous cell carcinoma
PIK3CA
siRNA
Proliferation
Migration
Apoptosis
