摘要
目的:探讨电转法沉默ATP结合盒转运子E1(ATP-binding cassette protein E1,ABCE1)基因的表达对人食管癌EC109细胞凋亡、增殖、侵袭及迁移的影响。方法:合成靶向ABCE1的siRNA序列(ABCE1-siRNA)以及阴性对照序列(NCsiRNA),电转法转染至EC109细胞,分别形成ABCE1-EC109、NC-siRNA-EC109细胞。RT-PCR、Western blotting检测转染后EC109细胞中ABCE1 mRNA与蛋白的表达情况,流式细胞术检测EC109细胞周期及凋亡,CCK-8法、划痕愈合实验、Transwell法分别检测EC109细胞的增殖、迁移以及侵袭的能力。结果:ABCE1-EC109细胞中ABCE1 mRNA和蛋白表达较NC-siRNAEC109细胞明显降低[(0.47±0.04)vs(0.67±0.05),(0.63±0.09)vs(0.86±0.11);均P<0.05]。与NC-siRNA-EC109细胞相比,ABCE1-EC109细胞的增殖速度明显减慢[(2.20±0.10)vs(2.91±0.13),P<0.05],细胞周期阻滞在G0/G1期细胞数目明显增多[(76.5±3.1)%vs(56.1±2.7)%,P<0.05)];细胞的凋亡率明显升高[(15.46±3.12)%vs(0.54±0.24)%,P<0.01],迁移、侵袭能力均显著下降[迁移:(8.12±0.23)vs(1.91±0.11)μm,P<0.05;侵袭:(42.56±4.68)vs(68.78±6.98)个,P<0.01]。结论:电转法沉默ABCE1基因的表达可促进食管癌EC109细胞的凋亡,抑制其体外增殖、侵袭及迁移。
Objective： To investigate the effect of silencing ATP-binding cassette protein E1 （ ABCE1） gene expression by electroporation on apoptosis, proliferation, migration and invasion of human esophageal squamous carcinoma EC-109 cells. Methods： The siRNA sequence （ABCE1-siRNA） targeting ABCE1 and the negative control sequence （NC-siRNA） were constructed and transfected into EC109 cells to obtain ABCE1-EC109 and NC-siRNA-EC109 cells, respectively. The expressions of ABCE1 mRNA and protein were detected by RT-PCR and Western blotting, respectively. Flow cytometry was used to detect the cell cycle and apoptosis of EC109 cells. The proliferation, migration and invasion of EC109 cells were evaluated by CCK-8 assay, wound closure assay and Transwell assay, respectively. Results： Compared with the NC-siRNA-EC109 cells, the expression levels of ABCE1 mRNA and protein were significantly decreased in the ABCE1-EC109 cells （／[0.47±0.04／] vs ／[0.67±0.05／], ／[0.63±0.09／] vs ／[0.86±0.11／], both P〈0.05）. Compared with the NC-siRNA-EC109 cells, the proliferation of ABCE1-EC109 cells was significantly decreased （／[2.20±0.10／] vs ／[2.91±0.13／]，P〈0.05）, the number of ABCE1-EC109 cells arrested at G0/G1 phase was increased （／[76.5±3.1／] vs ／[56.1±2.7／], P〈0.05）, the cell apoptotic rate was increased （／[15.46±3.12／] vs ／[0.54±0.24／], P〈0.01）, and the migration and invasion abilities were significantly decreased （migration： ／[8.12±0.23／] vs ／[1.91±0.11／], P〈005; invasion： ／[42.56±4.68／] vs ／[68.78±6.98／]，P〈0.01）. Conclusion： ABCE1 gene silencing by electroporation promotes the apoptosis of esophageal squamous carcinoma EC109 cells, and inhibits their proliferation, migration and invasion in vitro.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2013年第5期569-574,共6页
Chinese Journal of Cancer Biotherapy
关键词
ATP
结合盒转运子E1
食管癌
电转法
EC109细胞
凋亡
增殖
侵袭
迁移
ATP-binding cassette protein E 1 （ ABCE1）
esophageal carcinoma
electroporation
EC109 cell
apoptosis
proliferation
migration
invasion
