中国普通外科杂志2012，Vol.21Issue(3)：304-311,8.
shRNA稳定抑制XIAP和survivin表达对胰腺癌细胞生物学特征的影响
Influence of stable inhibition of XIAP and survivin expression on biological characteristics of pancreatic cancer cells
摘要
Abstract
Objective: To investigate the inhibitory effect of lentiviral vector-mediated RNA interference (RNAi) on XIAP (X-linked inhibitor of apoptosis protein) and survivin expression in human pancreatic cells and its effect on the proliferation, apoptosis and chemosensitivity of pancreatic cells.Methods: The pGCSIL-PUR and pGCSIL-NEO were used to construct the lentiviral vectors containing shRNA targeting XIAP and survivin gene, respectively, and the human pancreatic cancer SW1990 and Panc-1 cells were transfected with these lentiviruses. The cells were screened by puromycin and neomycin double selection and expanded to obtain the stably transfected cell lines. The XIAP and survivin expressions were determined by real-time PCR and Western blot, respectively. The proliferation and chemosensitivity of the cells were detected by MTT assay, and caspase-3/7 activity measurement and DAPI-staining were performed to assess cell apoptosis.Results: The stable SW1990 and Panc-1 cells with inhibition of XIAP and survivin expression were obtained after the effective interference targets of XIAP and survivin screening. MTT results showed that the proliferative abilities of both types of pancreatic cells significantly decreased after the suppression of XIAP or survivin expression, and the combination inhibition of the two genes exerted a strengthened effect. Although XIAP or survivin inhibition had no obvious effect on apoptosis of both types of pancreatic cells, they enhanced the sensitivities of both types of cells to the chemotherapy agents (5-FU and gemcitabine), and this effect was also strengthened by their combination.Conclusion: The lentiviral vector-mediated RNAi targeting XIAP and survivin can effectively inhibit XIAP and survivin expression, and also inhibit proliferation and enhance chemosensitivity of pancreatic cells. The combination inhibition of both genes can exert a synergistic effect.关键词
胰腺肿瘤/RNA干扰/X-连锁凋亡抑制蛋白/生存素/慢病毒Key words
Pancreatic Neoplasms/ RNA Interference/ X-linked inhibitor of apoptosis protein/ Survivin/ Lentivirus分类
医药卫生引用本文复制引用
宰红艳,江春,易小平,邓公平,李宜雄..shRNA稳定抑制XIAP和survivin表达对胰腺癌细胞生物学特征的影响[J].中国普通外科杂志,2012,21(3):304-311,8.基金项目
国家自然科学基金资助项目(30872492) （30872492）
湖南省自然科学基金资助项目(08JJ3042). （08JJ3042）
