摘要
目的:探讨PKA基因对儿童急性T淋巴细胞白血病的作用及其机制。方法:在RNA干扰技术沉默Jurkat细胞和Sup-T1细胞PKA基因后,采用CCK-8法、Transwell实验、细胞集落形成实验和流式细胞术研究下调PKA基因表达对Jurkat细胞和Sup-T1细胞增殖、迁移及细胞周期的作用。采用Western blot法检测转染si RNA后相关细胞周期蛋白水平。结果:转染针对PKA基因si RNA后,PKA基因在Jurkat细胞和Sup-T1细胞表达量均有不同程度的下降(P<0.05)。转染组的Jurkat细胞和Sup-T1细胞与对照组相比,其增殖较慢(P<0.05)。与阴性对照组的细胞相比,转染组Jurkat、Sup-T1细胞迁移和侵袭能力均明显减弱(P<0.05);同时细胞集落数量明显减少(P<0.05)。转染组的Jurkat细胞和Sup-T1细胞均明显阻滞在G0/G1期(P<0.05)。转染组的Jurkat和Sup-T1细胞中CDK2、Cyclin D1和p-Rb表达水平与对照组相比受到明显抑制(P<0.05)。结论:PKA基因表达下调可抑制急性T淋巴细胞白血病肿瘤细胞的活力,其分子机制可能与通过调控周期蛋白影响细胞分裂有关。
Objective： To explore the effect of PKA gene on acute T lymphocyte leukemia cells in children and its mechanism. Methods： Jurkat and Sup-T1 cells were divided into 2 group： control group（ Jurkat and Sup-T1 cells treated with non-specific si RNA） and transfected group（ Jurkat and Sup-T1 cells transfected with PKA si RNA）. The effects of down-regulating the expression of PKA gene on the viability,proliferotion,migration and cell cycle distribution of Jurkat and Sup-T1 cells in 2 groups were analyzed by CCK-8 assay,transwell experiment,cell colony-formation test and flowcytometry; the cyclin-related protein levels after transfection with PKA si RNA were detected by Western blot.Results： It was revealed that the expression of PKA in Jurkat and Sup-T1 cells decreased to different degree after si RNA transfection（ P〈0. 05）. CCK-8 assay showed that the proliferation of Jurkat and Sup-T1 cells in the transfected group was slower than that in the control group（ P〈0. 05）. Transwell experiment showed that the migration and invasion ability of Jurkat cells in the transfection group was weaker than that in the control group（ P〈0. 05）. The cell colong formation number of Jurkat and Sup-T1 cells in the transfection group decreased significantly（ P〈0. 05）. The cell level of Jurkat and Sup-T1 cells in G0/G1 phase increased after tansfection（ P〈0. 05）. Western blot assay revealed that the expression levels of CDK2,Cyclin D1 and p-Rb in the Jurkat and Sup-T1 cells of the transfection group were suppressed（ P〈0. 05）. Conclusion： The down-regulating PKA gene expression can decrease the proliferation and migration of tumor cells,and also can restrict the cell proliferation through related cell cycle proteins.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2018年第1期126-131,共6页
Journal of Experimental Hematology
