BCL11A-siRNA对HEK293细胞增殖的影响  

Influence of BCL11A-siRNA on HEK293 cell proliferation

作　　者：高杨军[1,2] 胡小毛[1] 吴红[1] 何冬梅[1] 赵军 李扬秋[1,3] 

机构地区：[1]暨南大学医学院血液病研究所,广州510632 [2]长治市人民医院 [3]暨南大学再生医学重点实验室

出　　处：《山东医药》2014年第3期4-7,共4页Shandong Medical Journal

基　　金：国家自然科学基金重大研究计划培育项目(91129720);国务院侨办重点学科建设基金资助项目(51205002);广东省科技计划重点项目(2009B0507000029)

摘　　要：目的设计靶向B细胞淋巴瘤/白血病11A基因的小干扰RNA(BCL11A-siRNA),并观察其对人胚胎肾细胞株HEK293细胞增殖的影响。方法利用siRNA Target Finder工具,针对BCL11A设计合成siRNA319、siRNA585、siRNA2292、siRNA475。将HEK293细胞接种不含抗生素的培养基中,分为阳性对照组(PC组)、阴性对照组(NC组)、BCL11A-siRNA319组(si319组)、BCL11A-siRNA585组(si585组)、BCL11A-siRNA2292组(si2292组)、BCL11A-siRNA475组(si475组)、空转组、细胞组。通过LipofectamineTM2000,si319组将siRNA319、si585组将siRNA585、si2292组将siRNA2292、si475组将siRNA475分别转染HEK293细胞;转染后6 h倒置荧光显微镜观察细胞转染情况并用流式细胞仪检测转染效率;转染后24、48、72 h,采用实时定量RT-PCR法观察BCL11A-siRNA对BCL11A基因表达的影响;CCK8法观察HEK293细胞体外增殖情况。结果转染后24 h,si585组、si2292组的HEK293细胞BCL11A基因表达降低,72 h逐渐恢复;si585组、si2292组细胞增殖受抑,与NC组、空转组、细胞组相比,P均<0.05。结论 BCL11AsiRNA585及BCL11A-siRNA2292可降低HEK293细胞中BCL11A基因表达,抑制细胞增殖。Objective To screen the effective small interfering RNAs （siRNAs） targeting against B cell lymphoma/ leukemia 11A（BCL11A） mRNA, and to observe the influence of BCL11A-siRNA on the proliferation of human embryonic kidney 293 （HEK 293） ceils. Methods Four siRNAs sequences targeting against BCLllA （siRNA, siRNA585, siR- NA2292, siRNA475） were designed and synthesized chemically by siRNA Target Finder. HEK293 cells were inoculated in the medium without antibiotics, and were divided into the positive control group （ PC group）, negative control group （ NC group）, BCL11A-siRNA319 group （ si319 group）, BCL11A-siRNA585 group （ si585 group）, BCL11A-siRNA2292 group （si2292 group）, BCL11A-siRNA475 group （si475 group）, idling group and cell group. Using lipofeetaminerM2000, all of the BCLllA-siRNAs （si319 group to siRNA319, si585 group to siRNA585, si2292 group to siRNA2292, si475 group to siRNA475 ） were transfected into HEK 293 cells. The transfection efficiency was detected by Flow eytometry, the BCL11A mRNA expression level was detected by real-time RT-PCR at 24, 48, 72 h after transfection and the cell proliferation was determined by CCK8 assay. Results At 24 h after transfection, the expression levels of BCLllA mRNA in si585 group, si2292 group were reduced, and recovery gradually at 72 h. The cell proliferation of the si585 group and si2292 group was inhibited as compared with the NC group, idling group and cell group （all P 〈 O. 05 ）. Conclusion BCL11A-siRNA585 and BCL11A-siRNA2292 could inhibit the expression of BCL11A gene and proliferation of HEK293 cells.

关 键 词：BCL11A基因 小干扰RNA HEK293细胞 细胞增殖 

分 类 号：R557.4[医药卫生—血液循环系统疾病]