Du Ning, Yang Bin, Hu Li-Juan, Zhao Yang, Sun Xin, Guo Zhi-Wei, Ren Hong
Department of Oncosurgery, First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an 710061, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2012 Apr;28(4):344-6.
To investigate the effects of Smac gene overexpression on chemotherapeutic sensitivity of esophageal cancer cell line Eca109 to cisplatin.
pcDNA3.1-Smac with GFP and pcDNA3.1-Smac with GFP were transfected into esophageal cancer cell line Eca109 by liposome and incubated with G418 for subclone selection.The efficiency of transfection was observed under fluorescence microscope, cellular Smac gene expression were determined by Western blot. Cisplatin treated group (1, 5, 10 mg/L) and cisplatin untreated group were selected to treat untransfected and transfected esophageal cancer cell line Eca109. Apoptosis was determined by Annexin V/PI.
The subclone esophageal cancer cell line Eca109, stable expressing Smac+GFP and neo+GFP respectively, were successfully selected, named as Eca109/Smac, Eca109/neo. Compared with the Eca109/neo and Eca109, the Smac expression level of Eca109/Smac was significantly increased(P<0.05). In cisplatin untreated group, the apoptosis rate was not associated with Smac expression. In cisplatin treated group (1, 5, 10 mg/L), compared with Eca109/neo and Eca109, the apoptosis rate of the Eca109/Smac was significantly increased after the treatments of cisplatin, the difference were significant(P<0.05). In addition, the apoptosis rate of Eca109/Smac was significantly increased with the concentration of cispaltin increased(P<0.05). The difference between Eca109/neo and Eca109 were not significant.
Smac gene didn't induce apoptosis in the cisplatin untreated Eca109 cells. Smac gene overexpression could increase chemotherapeutic sensitivity of esophageal cancer cell line Eca109 to cisplatin.
探讨Smac基因过表达对食管癌细胞系Eca109顺铂化疗敏感性的影响。
采用脂质体法将携带绿色荧光蛋白（GFP）的pcDNA3.1-Smac和携带GFP的pcDNA3.1-Smac转染至食管癌细胞系Eca109，并用G418进行亚克隆筛选。在荧光显微镜下观察转染效率，采用蛋白质免疫印迹法检测细胞Smac基因表达。选取顺铂处理组（1、5、10mg/L）和顺铂未处理组处理未转染及转染的食管癌细胞系Eca109，采用膜联蛋白V/碘化丙啶（Annexin V/PI）法检测细胞凋亡情况。
成功筛选出分别稳定表达Smac+GFP和neo+GFP的亚克隆食管癌细胞系Eca109，命名为Eca109/Smac、Eca109/neo。与Eca109/neo和Eca109相比，Eca109/Smac的Smac表达水平显著升高（P<0.05）。在顺铂未处理组，细胞凋亡率与Smac表达无关。在顺铂处理组（1、5、10mg/L），与Eca109/neo和Eca109相比，顺铂处理后Eca109/Smac的凋亡率显著升高，差异有统计学意义（P<0.05）。此外，随着顺铂浓度升高，Eca109/Smac的凋亡率显著升高（P<0.05）。Eca109/neo和Eca109之间差异无统计学意义。
Smac基因在顺铂未处理的Eca109细胞中不诱导凋亡。Smac基因过表达可提高食管癌细胞系Eca109对顺铂的化疗敏感性。
