bcl-2基因过表达致急性髓系白血病对柔红霉素耐药机制的初步探讨

发布时间：2019-03-10 20:43

【摘要】：目的探讨bcl-2基因过表达导致人急性髓系白血病对柔红霉素耐药的机制。方法通过分子克隆技术构建质粒pc DNA3.1(+)/bcl-2,将其转染人急性髓系白血病U937细胞,分正常对照组、空载体对照组、转染组,与柔红霉素培养24 h,采用CCK-8法检测细胞增殖率,Western blot检测凋亡相关蛋白的表达变化,从而比较转染bcl-2基因前后,U937细胞对柔红霉素敏感性的变化。结果成功构建质粒pc DNA3.1(+)/bcl-2,转染入U937细胞,PCR证实其转染效果;CCK-8法检测结果显示U937/bcl-2组细胞存活率显著高于对照组和空载体对照组;bcl-2基因过表达降低caspase-3片段的裂解活化,降低U937细胞对柔红霉素的敏感性。结论bcl-2过表达抑制caspase-3激活、抑制柔红霉素杀伤U937细胞,导致细胞对柔红霉素耐药。bcl-2可作为临床治疗急性髓系白血病的靶点。
[Abstract]:Objective to investigate the mechanism of daunorubicin (daunorubicin) resistance in acute myeloid leukemia induced by overexpression of bcl-2 gene. Methods Human acute myeloid leukemia cell line U937 was transfected with plasmid pc DNA3.1 () / bcl-2, by molecular cloning technique. The cells were divided into normal control group, empty vector control group, transfection group and daunorubicin culture for 24 h, and the cells were cultured with daunorubicin for 24 h. Cell proliferation rate, Western blot was used to detect the expression of apoptosis-related proteins in U937 cells. The sensitivity of U937 cells to daunorubicin (daunorubicin) was compared before and after bcl-2 gene transfection. Results the plasmid pc DNA3.1 () / bcl-2, was successfully constructed and transfected into U937 cells, and the transfection effect was confirmed by PCR. The cell survival rate of U937/bcl-2 group was significantly higher than that of control group and empty vector control group by CCK-8 assay. The overexpression of bcl-2 gene decreased the activation of caspase-3 fragment and decreased the sensitivity of U937 cells to daunorubicin. Conclusion the overexpression of bcl-2 inhibits the activation of caspase-3, inhibits daunorubicin killing U937 cells and leads to drug resistance to daunorubicin. Bcl-2 can be used as a target for clinical treatment of acute myeloid leukemia.
【作者单位】：南昌大学第一附属医院血液科;
【基金】：国家自然科学基金(编号:81200372) 江西省卫生计生委科技计划(编号:20165167)
【分类号】：R733.71

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