Department of Oncology, Affiliated Hospital, Guangdong Medical College, Zhanjiang, China.
Arch Med Sci. 2012 Dec 20;8(6):987-92. doi: 10.5114/aoms.2012.31251. Epub 2012 Oct 16.
To explore the effect of tumor suppressor in lung cancer 1 (TSLC1) on proliferation and apoptosis in esophageal cancer Eca109 cells.
Eca109 cells were divided into three groups: TSLC1 transfected group (TTG), mock group (MG) and untransfected group (UTG). The TTG and MG were transfected transiently with the pIRES2-EGFP-TSLC1 eukaryotic expression vector and pIRES2-EGFP vector respectively. The UTG was a blank control. The TSLC1 expression in TTG was analyzed with the fluorogram and RT-PCR method. Cell proliferation was measured with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) assay. Cell cycle was measured by flow cytometry (FCM). Cell apoptosis was detected by Annexin-V/PI double staining FCM.
Green color was found in TTG and MG. The band of TSLC1 mRNA of TTG was located at about 1400 bp by RT-PCR and agarose gel electrophoresis assay. The TSLC1 inhibited cell proliferation significantly in MTT assay, and the cell proliferation was slower in TTG than MG and UTG. After TSLC1 transfection, cell numbers increased in G0/G1 phase and decreased in S phase. Forty-eight hours after transfection, the apoptosis rate and death rate of TTG were higher than MG and UTG. Thus TSLC1 induced Eca109 cells to apoptosis.
The TSLC1 gene had a potent effect on cell proliferation inhibition, G1/S cell cycle arrest and induction of cell apoptosis in Eca109 cells.
探讨抑癌基因 1(TSLC1)对食管癌 Eca109 细胞增殖和凋亡的影响。
将 Eca109 细胞分为三组:TSLC1 转染组(TTG)、空载组(MG)和未转染组(UTG)。TTG 和 MG 分别用 pIRES2-EGFP-TSLC1 真核表达载体和 pIRES2-EGFP 载体瞬时转染。UTG 为空白对照。用荧光图和 RT-PCR 法分析 TTG 中 TSLC1 的表达。用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四唑(MTT)法测定细胞增殖。用流式细胞术(FCM)测定细胞周期。用 Annexin-V/PI 双染 FCM 检测细胞凋亡。
TTG 和 MG 均呈绿色。TTG 经 RT-PCR 和琼脂糖凝胶电泳检测,TSLC1mRNA 条带位于约 1400bp。MTT 法检测发现 TSLC1 明显抑制细胞增殖,TTG 组细胞增殖较 MG 组和 UTG 组缓慢。转染后,细胞 G0/G1 期数量增加,S 期数量减少。转染 48 小时后,TTG 的凋亡率和死亡率均高于 MG 和 UTG。因此,TSLC1 诱导 Eca109 细胞凋亡。
TSLC1 基因对 Eca109 细胞的增殖抑制、G1/S 细胞周期阻滞和诱导细胞凋亡有较强作用。