白血病·淋巴瘤
白血病·淋巴瘤
백혈병·림파류
JOURNAL OF LEUKEMIA & LYMPHOMA
2014年
6期
321-324,338
,共5页
于滕滕%贾玉娇%邱少伟%王厚才%邢海燕%田征%唐克晶%饶青%王敏
于滕滕%賈玉嬌%邱少偉%王厚纔%邢海燕%田徵%唐剋晶%饒青%王敏
우등등%가옥교%구소위%왕후재%형해연%전정%당극정%요청%왕민
RNA,小分子干扰%FHL2基因%K562细胞%细胞增殖%细胞凋亡
RNA,小分子榦擾%FHL2基因%K562細胞%細胞增殖%細胞凋亡
RNA,소분자간우%FHL2기인%K562세포%세포증식%세포조망
RNA,small interfering%FHL2%K562 cells%Cell proliferation%Apoptosis
目的 探讨FHL2基因对人类急性红白血病细胞生物学功能的影响.方法 应用Western blot检测常见白血病细胞系(K562、HEK293T、U937、HL-60、Kasumi-1、SKNO-1、NB-4及Nalm-6)FHL2蛋白表达情况.采用RNA干扰技术设计并合成FHL2 mRNA特异性shRNA,构建pLKO.1-puro干扰载体,制备慢病毒并感染人类急性红白血病K562细胞;应用实时定量聚合酶链反应(PCR)方法检测干扰效率;采用四甲基偶氮唑蓝(MTT)法检测干扰FHL2对细胞增殖能力的影响;用流式细胞术分析细胞凋亡的变化.结果 FHL2在K562、Kasumi-1和Nalm-6等白血病细胞系中表达,尤其在K562细胞中高表达.Western blot检测显示干扰FHL2的K562细胞FHL2蛋白相对表达水平较转染阴性对照序列的K562细胞明显下降(相对表达水平分别为1、0.284,t=8.872,P=0.0004),干扰K562细胞中FHL2基因表达能够抑制细胞的增殖,促进细胞的凋亡.结论 FHL2基因在多种白血病细胞系中表达升高.shRNA靶向干扰FHL2基因的表达可有效抑制K562细胞的增殖,促进其凋亡.
目的 探討FHL2基因對人類急性紅白血病細胞生物學功能的影響.方法 應用Western blot檢測常見白血病細胞繫(K562、HEK293T、U937、HL-60、Kasumi-1、SKNO-1、NB-4及Nalm-6)FHL2蛋白錶達情況.採用RNA榦擾技術設計併閤成FHL2 mRNA特異性shRNA,構建pLKO.1-puro榦擾載體,製備慢病毒併感染人類急性紅白血病K562細胞;應用實時定量聚閤酶鏈反應(PCR)方法檢測榦擾效率;採用四甲基偶氮唑藍(MTT)法檢測榦擾FHL2對細胞增殖能力的影響;用流式細胞術分析細胞凋亡的變化.結果 FHL2在K562、Kasumi-1和Nalm-6等白血病細胞繫中錶達,尤其在K562細胞中高錶達.Western blot檢測顯示榦擾FHL2的K562細胞FHL2蛋白相對錶達水平較轉染陰性對照序列的K562細胞明顯下降(相對錶達水平分彆為1、0.284,t=8.872,P=0.0004),榦擾K562細胞中FHL2基因錶達能夠抑製細胞的增殖,促進細胞的凋亡.結論 FHL2基因在多種白血病細胞繫中錶達升高.shRNA靶嚮榦擾FHL2基因的錶達可有效抑製K562細胞的增殖,促進其凋亡.
목적 탐토FHL2기인대인류급성홍백혈병세포생물학공능적영향.방법 응용Western blot검측상견백혈병세포계(K562、HEK293T、U937、HL-60、Kasumi-1、SKNO-1、NB-4급Nalm-6)FHL2단백표체정황.채용RNA간우기술설계병합성FHL2 mRNA특이성shRNA,구건pLKO.1-puro간우재체,제비만병독병감염인류급성홍백혈병K562세포;응용실시정량취합매련반응(PCR)방법검측간우효솔;채용사갑기우담서람(MTT)법검측간우FHL2대세포증식능력적영향;용류식세포술분석세포조망적변화.결과 FHL2재K562、Kasumi-1화Nalm-6등백혈병세포계중표체,우기재K562세포중고표체.Western blot검측현시간우FHL2적K562세포FHL2단백상대표체수평교전염음성대조서렬적K562세포명현하강(상대표체수평분별위1、0.284,t=8.872,P=0.0004),간우K562세포중FHL2기인표체능구억제세포적증식,촉진세포적조망.결론 FHL2기인재다충백혈병세포계중표체승고.shRNA파향간우FHL2기인적표체가유효억제K562세포적증식,촉진기조망.
Objective To investigate the influence of FHL2 gene on the biological behaviors of human acute leukemia cell line.Methods The expression of FHL2 gene in leukemic cells (K562,HEK293T,U937,HL-60,Kasumi-1,SKNO-1,NB-4 and Nalm-6) was assayed by Western blot.Lentviral vectors pLKO.1-puro containing the FHL2-shRNA were constructed and transfected into leukemic cells to downregulate FHL2 expression compared with the scramble shRNA,and to delineate its biological effects.The expression of FHL2 after shRNA treatment in the K562 cells was assayed by real-time quantitative PCR and Western blot.Effect of FHL2-shRNA on the proliferation of K562 cells was examined by MTT method.Apoptosis analysis was examined by FACS.Results FHL2 was expressed in K562,Kasumi-1 and Nalm-6 cell lines and so on.The expression level of FHL2 protein in K562 cells interfered by FHL-shRNA was lower than that in cells interfered by negtive control shRNA (1 vs 0.284,t =8.872,P =0.000 4).Down-regulation of FHL2 by shRNA in K562 cells led to the decreased cell proliferation and increased cell apoptosis.Conclusion The expression level of FHL2 is increased in many kinds of acute leukemia cell lines.The shRNA targeting FHL2 can effectively inhibit the proliferation and increase apoptosis of K562 cell line.
