CAJ | 학술논문

目的研究细胞黏附分子VLA-4抗体联合化疗药物依托泊苷(VP_(16))对儿童白血病细胞凋亡的影响,探讨骨髓基质细胞(BMSC)对儿童白血病细胞的保护作用及其可能的机制.方法分离儿童白血病骨髓单个核细胞,以BMSC+白血病细胞组为对照,在此基础上分别加入VLA4抗体和VP_(16)组成抗体组、药物组及抗体加药物组等三个实验组,采用流式细胞术检测各组白血病细胞的凋亡率;采用RT-PCR方法检测各组白血病细胞Survivin、bcl-2基因的表达.结果与对照组比较,三个实验组12 h及24 h的早期凋亡率及总凋亡率均显著增高(P<0.05);抗体加药物组白血病细胞早期凋亡率及总凋亡率最高(P<0.05);除对照组,三个实验组白血病细胞24 h的早期凋亡率及总凋亡率均显著高于12 h(P<0.01).三个实验组Survivin、bcl-2基因的表达均有不同程度的降低,抗体加药物组降低最明显.结论 BMSC对白血病细胞具有保护作用,VLA-4抗体能够抑制白血病BMSC与白血病细胞间的黏附,促进白血病细胞的凋亡,并且增强白血病细胞对VP_(16)诱导凋亡的敏感性.
목적 연구세포점부분자VLA-4항체연합화료약물의탁박감(VP_(16))대인동백혈병세포조망적영향,탐토골수기질세포(BMSC)대인동백혈병세포적보호작용급기가능적궤제.방법 분리인동백혈병골수단개핵세포,이BMSC+백혈병세포조위대조,재차기출상분별가입VLA4항체화VP_(16)조성항체조、약물조급항체가약물조등삼개실험조,채용류식세포술검측각조백혈병세포적조망솔;채용RT-PCR방법검측각조백혈병세포Survivin、bcl-2기인적표체.결과 여대조조비교,삼개실험조12 h급24 h적조기조망솔급총조망솔균현저증고(P<0.05);항체가약물조백혈병세포조기조망솔급총조망솔최고(P<0.05);제대조조,삼개실험조백혈병세포24 h적조기조망솔급총조망솔균현저고우12 h(P<0.01).삼개실험조Survivin、bcl-2기인적표체균유불동정도적강저,항체가약물조강저최명현.결론 BMSC대백혈병세포구유보호작용,VLA-4항체능구억제백혈병BMSC여백혈병세포간적점부,촉진백혈병세포적조망,병차증강백혈병세포대VP_(16)유도조망적민감성.
Objective To investigate the influence of very late antigen-4 (VLA-4) antibody and VLA-4 antibody combined with VP_(16) in vitro on childhood leukemic cells' apoptosis and explore the protection of bone marrow stromal cells (BMSC) upon leukemic cells and its related mechanisms. Methods Leukemic bone marrow stromal cells were isolated by human lymphocyte separation medium and in vitro culture of BMSC (adherent) and leukemia cells (suspended) BMSC+leukemic cells group were as control. Then VLA-4 antibody and/or VP_(16) were added respectively to VLA-4 antibody group, VP_(16) group and VLA-4 antibody combined with VP_(16) group to detect the apoptosis of leukemic cells in different groups through Annexin V-FITC double-labeled flow cytometry and the expression of Survivin, bcl-2 genes in each group of leukemic cells detected by RT-PCR. Results The results showed by flow cytometry that compared with the control groups, for 12 h or 24 h, the early and total apoptosis rates of leukemic cells of the three experimental groups were significantly increased(P <0.05); the early and total apoptosis rates of leukemic cells treated with VLA-4 antibody combined with VP_(16) group was markedly increased, compared with the control group (P <0.05); the comparison of the early and total apoptosis rates for the three experimental groups between 12 h and 24 h was significantly different (P<0.01). Moreover, RT-PCR results showed that the expression of Survivin and bcl-2 genes of leukemic cells in three experimental groups was reduced in varying degrees and the reduction of VLA-4 antibody combined with VP_(16) group was the most obvious. Conclusion BMSC plays a protective role on leukemic cells, and VLA-4 antibody can block the adhesion between BMSC and leukemic cells promoting leukemic cells apoptosis and enhance the sensibility of apoptosis of leukemic cells induced by chemotherapeutics.