Apollon siRNA抑制胰腺癌SW1990细胞增殖的研究

摘要

Objective To study the effect of small interfering RNA targeted on Apollon for proliferation on pancreatic cancer cells and its possible acting mechanism.Methods The small interfering RNA targeted on apollon in our previous study was transfected to the cells using LipofectamineTM 2000,after 48 hours transtection.The inhibitory effects of small interfering RNA targeted on Apollon (Apollon siRNA)on cell proliferation were detected by WST-8.Their inhibition rate and IC50 were calculated.The percentage of apoptosis cells were determined by flow cytometry.The expression of Apollon mRNA was analyzed by real time fluorescent quantitative reverse transcription polymerase chain reaction.The Apollon protein ex-pression levels were detected by western blotting.Results Apollon siRNA could effectively inhibit the proliferation of pan-creatic cancer cell.The amount of apoptotic cells increased significantly.The early apoptotic rate was 37.1%,and the RT-PCR results showed that the relative expression levels of Apollon mRNA were down-regulate,and shows a dose-effective-ness relations.The protein expression levels were decreased by Apollon siRNA.Conclusion Apollon siRNA can effectively inhibit the proliferation of pancreatic cancer cell.The mechanism may be work together to promote pancreative cancer cell early apoptosis and decreased the expression levels of gene and protein,which provides a novel potential approach for treat-ment of target therapy of pancreatic cancer.%目的：研究凋亡抑制蛋白（inhibitor of apoptosis proteins，IAPs）家族成员 Apollon小干扰 RNA（small interference RNA，siRNA）对人胰腺癌 SW1990细胞增殖的影响，并进一步探讨其作用机制。方法将前期研究的 Apollon siRNA的序列，经脂质体包裹转染胰腺癌（SW1990）细胞48 h后，采用WST-8法检测Apollon siRNA对胰腺癌SW1990细胞增殖抑制作用，并计算 IC50。流式细胞术检测细胞早期凋亡率；RT-PCR检测 Apollon mRNA的相对表达水平，western-blotting检测 Apollon蛋白的表达水平。结果 Apollon siRNA作用于胰腺癌（SW1990）细胞48 h后，能有效抑制 SW1990细胞增殖，并呈现剂量效应关系；流式细胞术检测结果显示：Apollon siRNA能促进胰腺癌（SW1990）细胞凋亡，其早期凋亡率达37.1％；RT-PCR结果显示，Apollon siRNA能显著下调Apollon mRNA及蛋白在胰腺癌细胞中的表达水平。结论 Apol-lon siRNA可抑制胰腺癌细胞的增殖，作用机制可能为共同促进胰腺细胞早期凋亡、下调 mRNA及蛋白的表达有关，为临床上胰腺癌的靶向基因治疗提供理论基础。