摘要
目的研究以bcl-2基因为靶标的短发夹RNA-3(shRNA-3)对急性原代白血病细胞存活和细胞对柔红霉素(DNR)敏感性的影响。方法将shRNA-3转入急性原代白血病细胞并与柔红霉素联合培养,于24、48和72h,用锥虫蓝拒染法计数活细胞,用免疫组织化学法检测白血病细胞bcl-2和P-糖蛋白(p-gp)的表达。结果shRNA-3能明显降低原代白血病细胞存活,以shRNA-3联合DNR作用最显著,在72h内活细胞数由2.0×105/mL下降到0.8×105/mL,并能显著抑制bcl-2和P-gP的表达,与单用DNR组相比,bcl-2蛋白表达率由(47.21±7.26)%下降到(27.82±8.25)%(P<0.05),P-gP表达率由(24.28±5.60)%下降到(16.34±4.15)%(P<0.05)。结论shRNA-3能增强急性原代白血细胞对柔红霉素的敏感性。
[Objective] To study the survive of leukemic cells and the effect of shRNA-3 targeting bel-2 gene on the drug-sensitivity of primary leukemic cell to Daunorubicin （DNR）. [Method] shRNA-3, which is leading sequence selected by previous experiments, was transferred into leukemic cell. Six hours later, the cells were cultured with DNR. The growth of leukemic cell was detected by Trypan blue dye exclusion test at 24, 48, 72 hours respectively. The level of bel-2 protein and P-gP was determined by immunoehemistry. [Results] shRNA-3 could significantly decline the survive of leukemic cells from acute leukemic, the co-euhrue of shRNA-3 with DNR was the most effective, in 72 hours, cell numbers from 2.0×10^5/mL down to 0.8×10^5/mL, and downregulate the expression of bcl-2 and P-gP oncogenes, hel-2 protein lowing rate from （47.21±7.26）% to （27.82±8.25）% （P 〈0.05）, and P-gP lowing rate from （24.28±5.60）% to （16.34±4.15）% （P 〈0.05）, as compared with alone of DNR. [Conclusion] The effective shRNA-3 targeting bel-2 ean enhanee the sensitivity of primary leukemic cell to DNR.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2009年第7期1056-1058,共3页
China Journal of Modern Medicine
基金
湖南省卫生厅资助项目(No:B20050102)
