PTTG反义cDNA对胆囊癌细胞5-FU敏感性的影响

PTTG antisense cDNA enhances sensitivity of gallbladder cancer cells to 5-FU

目的:探讨垂体肿瘤转化基因(PTTG)反义cDNA对胆囊癌GBC -SD细胞系5 -FU化疗敏感性的影响. 方法:脂质体法将PTTG反义cDNA转染胆囊癌细胞GBS- SD;G418筛选阳性克隆;Westernblot检测PTTG蛋白;MTT检测细胞增殖情况及不同浓度5- FU作用下各组细胞相对存活率差别,计算IC50值;流式细胞术检测各组细胞凋亡率. 结果:转染组细胞较未转染组PTTG表达明显减少;转染组细胞增殖与凋亡明显增强;转染组IC50值( 0. 605mmol/L)明显低于未转染组(1. 117mmol/L)和空载体转染组(0. 914mmol/L);转染组细胞的5 FU凋亡诱导作用( 26. 24% )明显高于未转染组(2. 67% )和空载体转染组(7. 62% ). 结论:转染PTTG反义cDNA可增强胆囊癌细胞对5- FU化疗敏感性,二者联合应用可能是一种治疗胆囊癌的有效方法.

AIM: To investigate the effect of PTTG antisense cDNA on the sensitivity of gallbladder cancer cells to 5-FU. METHODS: The recombinant vector containing PTTG antisense cDNA was transfected into GBC-SD gallbladder cancer cells. The positive cell clones were screened by G418. PTTG protein expression was detected by Western blot analysis. The changes of the cell proliferation and cell relative viability after treatment with different concentrations of 5-FU were assessed by MTT, the dose-effects curves were obtained and IC 50 s were calculated. Flow cytometry was used to demonstrate discrepancy of apoptotic rates. RESULTS: The expression of PTTG protein in transfected cells was decreased significantly as compared with that in non-transfected ones. The cell proliferation was accelerated and the cell apoptotic rate was higher in transfected cells. The IC 50 of 5-FU was significantly lower in transfected cells (0.605 mmol/L) than in non-transfected and empty vector transfected cells (1.117 mmol/L and 0.914 mmol/L respectively). After treatment with 5-FU, the apoptotic ratio was significantly higher in transfected cells (26.24%) than in non-transfected and empty vector transfectedcells (2.67% and 7.62% respectively). CONCLUSION: PTTG antisense cDNA transfection enhances the sensitivity of gallbladder cancer cells to 5-FU. Such an enhancement of sensitivity suggests that the combination of PTTG antisense cDNA with 5-FU can be an effective treatment modality for gallbladder cancer.