摘要
目的:探讨surv iv in反义核酸对阿霉素诱导胰腺癌细胞系PANC-1细胞凋亡的影响。方法:将已构建成功的surv iv in反义核酸真核表达载体pcDNA 3-SVV as电转染胰腺癌细胞系PANC-1,并筛选转染成功的阳性克隆;台盼蓝拒染法观察surv iv in反义核酸与阿霉素联合应用对PANC-1细胞生存的影响;细胞计数和M TT试验测定转染细胞对阿霉素敏感性;琼脂糖凝胶电泳分析细胞凋亡DNA断裂情况。结果:转染surv iv in反义核酸的PANC-1/SVV as细胞增殖明显受抑,与PANC-1/neo细胞、PANC-1细胞进行比较,具有显著性差异(P<0.05);M TT实验显示,阿霉素对PANC-1/SVV as、PANC-1/neo、PANC-1细胞的IC50值分别为0.285±0.012μm o l/L、1.528±0.317μm o l/L和1.540±0.253μm o l/L。统计分析证明差异具有显著性(P<0.01);经琼脂糖凝胶电泳,PANC-1/SVV as细胞可见到DNA梯形条带,而PANC-1/neo、PANC-1细胞未见到。结论:surv iv in反义核酸可促进阿霉素诱导PANC-1细胞凋亡,为胰腺癌的基因治疗研究提供了实验依据。
Objective： To explore the effects of survivin antisense RNA on doxorubicin - induced apoptosis in pancreatic cancer cell line PANC-1. Methods： A survivin antisense eukaryotic vector pcDNA3- SVVas prepared in previous study was delivered into PANC- 1 mediated by electroperforation. Cell survival fraction was determined using the trypan blue dye exclusion assay. Cell proliferation and MTT assay were tested to investigate the sensibility of transfected cells to doxorubicin. Apoptosis was detected by DNA gel electrophoresis. Results： We obtained two positive cell clone PANC - 1/SVVas and PANC- 1/neo by eletroporation. Compared to PANC- 1 and PANC- 1/neo cells, PANC- 1/SVVas cells growth was significantly reduced （ P 〈0. 05）. By MTT assay, the IC50 of to PANC- 1/SVVas, PANC- 1/neo and PANC-1 cells were 0. 285±0. 012μmol/L, 1. 528±0. 317μmol/L and 1. 540±0. 253μmol/L respectively, the difference was significant by statistic analysis （ P 0. 01）. Agarose gel electrophoresis of genomic DNA from PANC-1/SVVas showed typical DNA ladder, but DNA from PANC-1/neo and PANC-1 did not. Conclusion： Survivin antisense RNA could enhance doxorubicin -induced apoptosis in pancreatic cancer cell line PANC-1. This may lay an experimental foundation for further research of gene therapy in pancreatic cancer.
出处
《内蒙古医学院学报》
2005年第3期180-184,共5页
Acta Academiae Medicinae Neimongol
基金
内蒙古自然科学基金(200308020605)
