RNAi沉默CDX2基因对急性髓细胞白血病细胞增殖影响的实验研究  

Effects of RNA interference to CDX2 gene on proliferation of human AML cell

作　　者：王东宁[1] 黎国伟[2] 叶玉蝶[2] 吴红梅[2] 许先吟[2] 陈立[2] 余相[2] 

机构地区：[1]中山大学附属第三医院血液科,广东广州510630 [2]惠州市中心人民医院血液科,广东惠州516001

出　　处：《中华肿瘤防治杂志》2010年第22期1811-1814,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：广东省医学科学研究基金(A2008751)

摘　　要：目的：探讨RNA干扰技术沉默CDX2基因对人急性髓细胞白血病细胞系THP1增殖影响的意义。方法：针对CDX2mRNA序列设计合成2对编码小干扰RNA（siRNA）的DNA模板,构建pGenSil-CDX2siRNA重组质粒,转染THP1细胞。采用蛋白质印迹法检测重组质粒对CDX2蛋白表达的影响,MTT法观察THP1细胞增殖情况,Annexin-V-PI双染法流式细胞术检测转染重组质粒后细胞凋亡状况。结果：成功构建pGenSil-CDX2 siRNA重组质粒,成功转染THP1细胞,并能特异地抑制CDX2基因的表达;转染重组质粒后,THP1的活力分别降低为（39.8±5.0）%,与空质粒组组间差异有统计学意义,P=0.016。重组质粒组的THP1细胞凋亡率为37.3%~42.1%。结论：pGenSil-CDX2 siRNA重组质粒明显下调CDX2在急性髓细胞白血病细胞中的表达,并抑制肿瘤细胞生长,促进其凋亡。OBJECTIVE：To study the effects of RNA interference to CDX2 gene on proliferation of human AML cell.METHODS：A eukaryotic expression vector pGenSil was used to carry out siRNA.The DNA inserts were synthesized to target CDX2 gene.By cloning technique,pGenSil-CDX2 siRNA was constructed and confirmed by restriction cutting.AML cell（THP1 cell） was used for experiment.The expression of CDX2 was detected by Wester blot;MTT and FCM were used to observe the proliferation and apoptosis of THP1 cells.RESULTS：pGenSil-CDX2 siRNA was constructed successfully.Western blot analyses found that the expression of the CDX2 protein in THP1 cells transfected with recombinant plasmid pGenSil-CDX2 siRNA decreased;MTT showed that it could suppress the growth of THP1 cells to（39.8±5.0）%,and there was significant difference compared with blank plasmid group（P=0.016）;FCM demonstrated that recombinant plasmid induced apoptosis of the cells 37.3%-42.1%.CONCLUSION：pGenSil-CDX2 siRNA can significantly inhibit CDX2 expression,suppress the growth of THP1 cells and induce apoptosis of the cells.

关 键 词：白血病 髓样 急性/病理学 转录因子 基因 细胞凋亡 

分 类 号：R733.71[医药卫生—肿瘤]