医学临床研究
醫學臨床研究
의학림상연구
JOURNAL OF CLINICAL RESEARCH
2013年
9期
1665-1667,1668
,共4页
白血病%RNA%单核细胞
白血病%RNA%單覈細胞
백혈병%RNA%단핵세포
Leukemia%RNA%Monocytes
[目的]探讨 Ets2基因在白血病中的表达情况及靶向小 RNA 片段（siRNA）沉默人单核细胞白血病细胞株（SHI-1）中 Ets2基因表达对 SHI-1化疗敏感性的影响。[方法]应用 RT-PCR 技术检测28例白血病患者骨髓标本和 SHI-1细胞中 Ets2表达水平。设计和体外化学合成特异性针对 Ets2基因的 siRNA；用电穿孔方法转染 siRNA 至 SHI-1细胞株后，QRT-PCR 检测 siRNA 对 Ets2基因 mRNA 表达的抑制效应；An-nexin V-FITC/PI 双染色法分别检测细胞凋亡率和靶向 siRNA 沉默 Ets2基因后对 VP-16诱导 SHI-1细胞凋亡率，并进行对比。[结果]Ets2基因 在 SHI-1细胞中高表达；Ets2表达水平在28例白血病骨髓标本中（71．4％）明显高于对照组（40％）（P ＜0．05）。siRNA 转染 SHI-1细胞后，细胞凋亡率增加，Ets2基因沉默后的 SHI-1细胞对依托泊苷（VP-16）化疗敏感性增强（P ＜0．01）。[结论]Ets2转录因子在白血病细胞中高表达可能与白血病细胞的发生发展有关。靶向 siRNA 通过沉默 Ets2表达可而增加 SHI-1细胞的凋亡和化疗敏感性，推测 Ets2高表达可能与白血病细胞对化疗药物的抵抗存在相关。
[目的]探討 Ets2基因在白血病中的錶達情況及靶嚮小 RNA 片段（siRNA）沉默人單覈細胞白血病細胞株（SHI-1）中 Ets2基因錶達對 SHI-1化療敏感性的影響。[方法]應用 RT-PCR 技術檢測28例白血病患者骨髓標本和 SHI-1細胞中 Ets2錶達水平。設計和體外化學閤成特異性針對 Ets2基因的 siRNA；用電穿孔方法轉染 siRNA 至 SHI-1細胞株後，QRT-PCR 檢測 siRNA 對 Ets2基因 mRNA 錶達的抑製效應；An-nexin V-FITC/PI 雙染色法分彆檢測細胞凋亡率和靶嚮 siRNA 沉默 Ets2基因後對 VP-16誘導 SHI-1細胞凋亡率，併進行對比。[結果]Ets2基因 在 SHI-1細胞中高錶達；Ets2錶達水平在28例白血病骨髓標本中（71．4％）明顯高于對照組（40％）（P ＜0．05）。siRNA 轉染 SHI-1細胞後，細胞凋亡率增加，Ets2基因沉默後的 SHI-1細胞對依託泊苷（VP-16）化療敏感性增彊（P ＜0．01）。[結論]Ets2轉錄因子在白血病細胞中高錶達可能與白血病細胞的髮生髮展有關。靶嚮 siRNA 通過沉默 Ets2錶達可而增加 SHI-1細胞的凋亡和化療敏感性，推測 Ets2高錶達可能與白血病細胞對化療藥物的牴抗存在相關。
[목적]탐토 Ets2기인재백혈병중적표체정황급파향소 RNA 편단（siRNA）침묵인단핵세포백혈병세포주（SHI-1）중 Ets2기인표체대 SHI-1화료민감성적영향。[방법]응용 RT-PCR 기술검측28례백혈병환자골수표본화 SHI-1세포중 Ets2표체수평。설계화체외화학합성특이성침대 Ets2기인적 siRNA；용전천공방법전염 siRNA 지 SHI-1세포주후，QRT-PCR 검측 siRNA 대 Ets2기인 mRNA 표체적억제효응；An-nexin V-FITC/PI 쌍염색법분별검측세포조망솔화파향 siRNA 침묵 Ets2기인후대 VP-16유도 SHI-1세포조망솔，병진행대비。[결과]Ets2기인 재 SHI-1세포중고표체；Ets2표체수평재28례백혈병골수표본중（71．4％）명현고우대조조（40％）（P ＜0．05）。siRNA 전염 SHI-1세포후，세포조망솔증가，Ets2기인침묵후적 SHI-1세포대의탁박감（VP-16）화료민감성증강（P ＜0．01）。[결론]Ets2전록인자재백혈병세포중고표체가능여백혈병세포적발생발전유관。파향 siRNA 통과침묵 Ets2표체가이증가 SHI-1세포적조망화화료민감성，추측 Ets2고표체가능여백혈병세포대화료약물적저항존재상관。
[Objective]To explore the expression of Ets2 gene and the influence of siRNA targeting Ets2 gene in human leukemia monocytic cells(SHI-1)on chemotherapy sensitivity of SHI-1.[Methods]Reverse transcription polymerase chain reaction(RT-PCR)was used to detect the expression of Ets2 gene in bone mar-row samples of 28 patients with leukemia and SHI-1 cells.The siRNA specific targeting Ets2 gene was de-signed and synthesized in vitro.After siRNA was transfected to SHI-1 cells by using electroporation method, the inhibition effect of siRNA on mRNA expression of Ets2 gene was detected by QRT-PCT.Annexin V-FITC/PI double staining method was used to detect cell apoptosis and etoposide(VP-16)-induced apoptosis of SHI-1 after siRNA targeting to Ets2 gene.[Results]The expression of Ets2 gene was high in SHI-1 cells. The expression of Ets2 in bone marrow samples of 28 patients with leukemia was 71.4%,which was obviously higher than that in control group(40%)(P <0.05).After siRNA was transfected to SHI-1 cells,cell apopto-sis rate increased.After siRNA targeting to Ets2 gene,VP-16 sensitivity of SHI-1 cells increased(P <0.01).[Conclusion]High expression of Ets2 transcription factor in leukemia cells may be associated with the patho-genesis and development of leukemia cells.The siRNA targeting Ets2 can increase the cell apoptosis and chem-otherapy sensitivity of SHI-1 cells.It supposes that high expression of Ets2 may be related to the chemothera-py drug resistance of leukemia.
