白血病·淋巴瘤
白血病·淋巴瘤
백혈병·림파류
Journal of Leukemia & Lymphoma
2015年
9期
535-538
,共4页
张艳君%贾秀红%李建厂%穆旭光
張豔君%賈秀紅%李建廠%穆旭光
장염군%가수홍%리건엄%목욱광
同源盒A10基因%多药耐药1基因%慢病毒%RNA干扰%NB4细胞
同源盒A10基因%多藥耐藥1基因%慢病毒%RNA榦擾%NB4細胞
동원합A10기인%다약내약1기인%만병독%RNA간우%NB4세포
HOXA10 gene%Multi-drug resistance-1 gene%Lentivirus%RNA interference%NB4 cell line
目的 探讨慢病毒载体介导的短发夹RNA(shRNA)靶向沉默同源盒A10(HOXA10)基因对白血病NB4细胞增殖、凋亡和耐药的影响.方法 将NB4细胞分为干扰(lenti-shHOXA10)组、阴性对照组和未处理组,用慢病毒感染NB4细胞5d后进行相关实验.运用流式细胞仪测定慢病毒对NB4细胞的感染效率.实时荧光聚合酶链反应(RT-PCR)、蛋白质印迹法(Western blot)测定慢病毒干扰载体对HOXA 10基因的沉默作用;MTF法检测细胞的增殖抑制情况.流式细胞术检测3组细胞凋亡率的变化,Western blot方法测定干扰HOXA10基因对多药耐药1(MDR-1)蛋白的影响.结果 慢病毒对NB4细胞的感染率达90％左右.干扰组HOXA10 mRNA和蛋白的表达水平均较对照组下降,干扰组细胞抑制率和凋亡率分别为(52.12±4.02)％、(30.0±2.7)％,与阴性对照组和未处理组相比差异均有统计学意义(P＜0.05).Western blot结果显示干扰HOXA 10基因的表达能降低MDR-1基因的表达水平,逆转白血病细胞耐药.结论 慢病毒载体靶向干扰HOXA10基因的表达,能有效抑制NB4细胞增殖和促进其凋亡,逆转白血病细胞耐药.HOXA10基因有望成为逆转白血病耐药的新靶点.
目的 探討慢病毒載體介導的短髮夾RNA(shRNA)靶嚮沉默同源盒A10(HOXA10)基因對白血病NB4細胞增殖、凋亡和耐藥的影響.方法 將NB4細胞分為榦擾(lenti-shHOXA10)組、陰性對照組和未處理組,用慢病毒感染NB4細胞5d後進行相關實驗.運用流式細胞儀測定慢病毒對NB4細胞的感染效率.實時熒光聚閤酶鏈反應(RT-PCR)、蛋白質印跡法(Western blot)測定慢病毒榦擾載體對HOXA 10基因的沉默作用;MTF法檢測細胞的增殖抑製情況.流式細胞術檢測3組細胞凋亡率的變化,Western blot方法測定榦擾HOXA10基因對多藥耐藥1(MDR-1)蛋白的影響.結果 慢病毒對NB4細胞的感染率達90％左右.榦擾組HOXA10 mRNA和蛋白的錶達水平均較對照組下降,榦擾組細胞抑製率和凋亡率分彆為(52.12±4.02)％、(30.0±2.7)％,與陰性對照組和未處理組相比差異均有統計學意義(P＜0.05).Western blot結果顯示榦擾HOXA 10基因的錶達能降低MDR-1基因的錶達水平,逆轉白血病細胞耐藥.結論 慢病毒載體靶嚮榦擾HOXA10基因的錶達,能有效抑製NB4細胞增殖和促進其凋亡,逆轉白血病細胞耐藥.HOXA10基因有望成為逆轉白血病耐藥的新靶點.
목적 탐토만병독재체개도적단발협RNA(shRNA)파향침묵동원합A10(HOXA10)기인대백혈병NB4세포증식、조망화내약적영향.방법 장NB4세포분위간우(lenti-shHOXA10)조、음성대조조화미처리조,용만병독감염NB4세포5d후진행상관실험.운용류식세포의측정만병독대NB4세포적감염효솔.실시형광취합매련반응(RT-PCR)、단백질인적법(Western blot)측정만병독간우재체대HOXA 10기인적침묵작용;MTF법검측세포적증식억제정황.류식세포술검측3조세포조망솔적변화,Western blot방법측정간우HOXA10기인대다약내약1(MDR-1)단백적영향.결과 만병독대NB4세포적감염솔체90％좌우.간우조HOXA10 mRNA화단백적표체수평균교대조조하강,간우조세포억제솔화조망솔분별위(52.12±4.02)％、(30.0±2.7)％,여음성대조조화미처리조상비차이균유통계학의의(P＜0.05).Western blot결과현시간우HOXA 10기인적표체능강저MDR-1기인적표체수평,역전백혈병세포내약.결론 만병독재체파향간우HOXA10기인적표체,능유효억제NB4세포증식화촉진기조망,역전백혈병세포내약.HOXA10기인유망성위역전백혈병내약적신파점.
Objective To explore the effects of lentivirus-mediated RNA interference targeting HOXA10 gene on the proliferation, apoptosis and drug resistance of leukemic cell line NB4.Methods NB4 cells were divided into three groups: interference group, negative control group and untreated group.The infection efficiency of lentivirus for NB4 cells was detected by flow cytometry, and the expression of HOXA10 gene of NB4 cells at mRNA and protein level was detected by real-time PCR and Western blot.Cell survival was determined by MTF assay, and apoptosis and necrosis rates were detected by flow cytometry.Western blot was used to detect the influence of down-regulation HOXA10 gene on the multi-drug resistance-1 (MDR-1) protein.Results The ratio of GFP positive cells was up to 90 ％.HOXA10 gene mRNA and protein levels were decreased in interference group compared with control group.The inhibition rate of interference group was (52.12±4.02) ％, the apoptosis rate of interference group was (30.0±2.7) ％, and their differences in the interference group and in control groups (negative control group and untreated group) were significant (P ＜ 0.05).Western blot results showed that interfering HOXA10 gene significantly reduced the resistance gene MDR-1 expression level and reverse the drug-resistant of leukemia cells.Conclusions Lentivirns-shHOXA10 can steadily reduce the expression level of HOXA10, inhibit the leukemic cells proliferation, promote apoptosis and reverse drug-resistant.HOXA10 gene is expected to become a new target for reversing leukemia drug resistance.
