摘要
目的探讨凋亡抑制蛋白Survivin反义寡核苷酸转染对胰腺癌BxPC 3细胞系SurvivinmRNA的表达及细胞增殖、凋亡和对吉西他滨化疗敏感性的影响。方法用脂质体瞬时转染法介导Survivin反义硫代磷酸寡核苷酸处理胰腺癌BxPC 3细胞后用RT PCR检测SurvivinmRNA的表达 ,四唑氮蓝法检测细胞的相对存活率 ,用流式细胞仪和透射电镜检测其对BxPC 3细胞的凋亡诱导作用。结果Survivin反义寡核苷酸作用于BxPC 3细胞后 ,细胞的存活率呈剂量和时间依赖性 ,其中 2 4h的IC50 值为 4 0 0nmol/L ,在此浓度和时间下 ,Survivin反义寡核苷酸可明显降低SurvivinmR NA的表达 ,细胞凋亡率为 (2 5± 3) %。在透射电镜下BxPC 3细胞可呈凋亡的早期改变。Survivin反义寡核苷酸和吉西他滨的联合实验组与单独应用吉西他滨组相比 ,在 4 8h和 72h可使细胞的存活率分别降低 2 76和 4 5 8倍。结论Survivin反义寡核苷酸可诱导胰腺癌细胞凋亡、抑制细胞增殖并增强吉西他滨的化疗敏感性。
Objective In this study,we determine whether Survivin antisense oligonucleotide (ASODN) down-regulates the expression of Survivin mRNA,induces apoptosis,and enhances the sensitivity of pancreatic cancer cells to a chemotherapy agent Gemcitabine. Methods Lipofectin was used to encapsulate ASODN in transfection. Viability of pancreatic cell line BxPC-3 cells treated with ASODN was assessed by MTT method,the expressions of Survivin mRNA were detected by RT-PCR;Flow cytometry and electron microscopy were used to demonstrate apoptotic changes in ASODN treated cells. Result Cell viability was inhibited in dose-dependent and time-dependent manner with an IC 50 of 400 nM at the time of 24 h,Survivin mRNA was down-regulated by 3.38 fold compared to control group. The cell apoptotic ratio was 24.93%±2.97%,early apoptotic morphology was demonstrated by electron microscopy. In combination with Gemcitabine,at the time of 48 h and 72 h,cell viability decreased by 2.76 and 4.58 fold compared to when Gemcitabine was used alone. Conclusion Survivin ASODN down-regulates Survivin mRNA ,induces apoptosis,inhibits proliferation and enhances the sensitivity of pancreatic cancer cells to Gemcitabine.
出处
《中华普通外科杂志》
CSCD
北大核心
2004年第7期401-403,共3页
Chinese Journal of General Surgery