白血病·淋巴瘤
白血病·淋巴瘤
백혈병·림파류
JOURNAL OF LEUKEMIA & LYMPHOMA
2012年
8期
453-455
,共3页
miRNA-214%反义核酸%U937细胞%细胞凋亡
miRNA-214%反義覈痠%U937細胞%細胞凋亡
miRNA-214%반의핵산%U937세포%세포조망
miRNA-214%Antisense oligonucleotides%U937 cells%Apoptosis
目的 探讨以miRNA-214为靶点的反义核酸对白血病U937细胞的生长抑制作用及可能的作用机制.方法 人工合成miRNA-214的反义核酸序列,硫代修饰,以Lipofectamine 2000为介导转入U937细胞.MTT细胞毒性检测U937细胞的增殖活性,流式细胞术检测细胞的凋亡水平,荧光定量PCR 检测miRNA-214的表达水平.结果 MTT检测结果显示,转染反义核酸后24、48、72 h,U937细胞的增殖活性显著下降(0.812±0.001、0.770±0.002、0.541±0.001),与随机对照组(1.011±0.002、1.112±0.003、1.111 ±0.003)、空白对照组(1.112±0.001、1.023±0.001、1.101±0.001)相比较差异均有统计学意义( F=2.782、3.659、2.735,P=0.021、0.018、0.036).流式细胞术检测结果表明,在转染反义核酸48 h后细胞的凋亡水平随时间延长而提高(48、72 h分别为15.12±0.02、19.14±0.01),与随机对照组(2.04±0.02、2.45±0.03)、空白对照组(1.19±0.02、2.02±0.01)相比较差异均有统计学意义(F=3.683、3.762,P=0.013、0.015).荧光定量PCR结果证实,在反义核酸作用后,U937细胞内miRNA-214的相对表达水平明显下降(31.1±0.2),与随机对照组、空白对照组的25.8±0.1、25.6±0.2相比较,差异均有统计学意义(均P＜ 0.05).结论 以miRNA-214为靶点的反义核酸可抑制U937细胞的增殖活性,并明显促进细胞的凋亡.
目的 探討以miRNA-214為靶點的反義覈痠對白血病U937細胞的生長抑製作用及可能的作用機製.方法 人工閤成miRNA-214的反義覈痠序列,硫代脩飾,以Lipofectamine 2000為介導轉入U937細胞.MTT細胞毒性檢測U937細胞的增殖活性,流式細胞術檢測細胞的凋亡水平,熒光定量PCR 檢測miRNA-214的錶達水平.結果 MTT檢測結果顯示,轉染反義覈痠後24、48、72 h,U937細胞的增殖活性顯著下降(0.812±0.001、0.770±0.002、0.541±0.001),與隨機對照組(1.011±0.002、1.112±0.003、1.111 ±0.003)、空白對照組(1.112±0.001、1.023±0.001、1.101±0.001)相比較差異均有統計學意義( F=2.782、3.659、2.735,P=0.021、0.018、0.036).流式細胞術檢測結果錶明,在轉染反義覈痠48 h後細胞的凋亡水平隨時間延長而提高(48、72 h分彆為15.12±0.02、19.14±0.01),與隨機對照組(2.04±0.02、2.45±0.03)、空白對照組(1.19±0.02、2.02±0.01)相比較差異均有統計學意義(F=3.683、3.762,P=0.013、0.015).熒光定量PCR結果證實,在反義覈痠作用後,U937細胞內miRNA-214的相對錶達水平明顯下降(31.1±0.2),與隨機對照組、空白對照組的25.8±0.1、25.6±0.2相比較,差異均有統計學意義(均P＜ 0.05).結論 以miRNA-214為靶點的反義覈痠可抑製U937細胞的增殖活性,併明顯促進細胞的凋亡.
목적 탐토이miRNA-214위파점적반의핵산대백혈병U937세포적생장억제작용급가능적작용궤제.방법 인공합성miRNA-214적반의핵산서렬,류대수식,이Lipofectamine 2000위개도전입U937세포.MTT세포독성검측U937세포적증식활성,류식세포술검측세포적조망수평,형광정량PCR 검측miRNA-214적표체수평.결과 MTT검측결과현시,전염반의핵산후24、48、72 h,U937세포적증식활성현저하강(0.812±0.001、0.770±0.002、0.541±0.001),여수궤대조조(1.011±0.002、1.112±0.003、1.111 ±0.003)、공백대조조(1.112±0.001、1.023±0.001、1.101±0.001)상비교차이균유통계학의의( F=2.782、3.659、2.735,P=0.021、0.018、0.036).류식세포술검측결과표명,재전염반의핵산48 h후세포적조망수평수시간연장이제고(48、72 h분별위15.12±0.02、19.14±0.01),여수궤대조조(2.04±0.02、2.45±0.03)、공백대조조(1.19±0.02、2.02±0.01)상비교차이균유통계학의의(F=3.683、3.762,P=0.013、0.015).형광정량PCR결과증실,재반의핵산작용후,U937세포내miRNA-214적상대표체수평명현하강(31.1±0.2),여수궤대조조、공백대조조적25.8±0.1、25.6±0.2상비교,차이균유통계학의의(균P＜ 0.05).결론 이miRNA-214위파점적반의핵산가억제U937세포적증식활성,병명현촉진세포적조망.
Objective To explore the inhibitory effect of anti-miRNA-214 oligonucleotide on leukemia U937 cells.Methods U937 cells were transfected with anit-miRNA-214 oligonucleotide,cell viability was analyzed by MTT assay.Apoptosis was detected by flow cytometry.The expression of miRNA214 in the U937 cells were measured by real-time PCR.Results The MTT result showed that the growth of U937 cells treated with AMO-miRNA-214 in 24 h,48 h,72 h was obviously inhibited (0.812±0.001,0.770±0.002,0.541±0.001),compared with those in control groups (randomized control 1.011±0.002,1.112±0.003,1.111±0.003,blank control 1.112±0.001,1.023±0.001,1.101±0.001),the differences had statistical significance (F =2.782,3.659,2.735,P =0.021,0.018,0.036).The flow cytometry results showed that the apoptosis detected in AMO-miRNA-214 group at 48 h,72 h (15.12±0.02,19.14±0.01) had significant differences compared with those in control groups (randomized control 2.04±0.02,2.45±0.03,blank control 1.19±0.02,2.02±0.01) (F =3.683,3.762,P =0.013,0.015).The expression of miRNA-214 was downregulated significantly in U937 cells after treated with oligonucleotide (31.1±0.2) compared with those in control groups (randomized control 25.8±0.1,blank control 25.6±0.2) (P ＜ 0.05).Conclusion Targeted inhibition of miRNA-214 with oligonucleotide can suppress U937 cells growth and induce apoptosis.
