骨髓基质细胞及抗晚期抗原4抗体联合阿糖胞苷对急性髓系白血病HL-60细胞增殖和凋亡的影响

Effects of bone marrow stromal cells and antibody against late antigen-4 combined with Ara-C on proliferation and apoptosis of HL-60 cells

目的探讨骨髓基质细胞、抗晚期抗原4抗体（aVLA-4）、阿糖胞苷（Ara-C）对白血病HL-60细胞增殖和凋亡的影响。方法实验共分为5组：HL-60细胞单独培养组（对照组），HL-60细胞＋基质细胞组（基质细胞组），HL-60细胞＋基质细胞＋aVLA-4组（抗体组），HL-60细胞＋基质细胞＋Ara-C组（药物组），HL-60细胞＋基质细胞＋aVLA-5＋Ara-C组（抗体＋药物组）。采用CCK-8法检测各组细胞增殖或抑制率，流式细胞术检测各组HL-60细胞凋亡情况，Western blot法检测各组HL-60细胞中抗凋亡基因bcl-2表达情况。结果基质细胞组培养24h及48h后HL-60细胞数量均高于对照组，差异有统计学意义[（7.2±0-3）×10^5／ml比（5.3±0.4）×10^5／ml，（8.4±0.2）×10^5／ml比（6.8±0.3）×10^5／ml（P〈0.001）]；与基质细胞组比较，抗体组、药物组、抗体＋药物组HL-60白血病细胞的增殖抑制率[（24_3±2.1）％、（37.0±2.6）％、（65.6±3.8）％]及凋亡率[（5.7±O.6）％、（8.0±0.5）％、（10.44±0.9）％、（16.5±0.7）％]均增高，差异有统计学意义（P〈0.05），并以抗体＋药物组的增高最为明显；各组的bcl-2蛋白表达均有下降，以抗体＋药物组的下降最明显。结论骨髓基质细胞对白血病细胞的增殖起促进作用，aVLA-4干扰基质细胞与白血病细胞间的相互作用可增强白血病细胞对Ara-C的化疗敏感性。

Objective To explore the bone marrow stromal cells, anti-late antigen-4 （VLA-4） antibody （aVLA-4）, cytarabine （Ara-C） on the proliferation and apoptosis of leukemia HL-60 cells. Methods The experiment was divided into five groups： HL-60 cells were cultured alone （control group）, HL-60 ceils and stromal cells group （stromal cells group）, HL-60 cells ＋ stromal cells ＋ aVLA-4 （antibody group）, HL-60 cells ＋ stromal cells ＋ Ara-C group （drug group）, HL-60 cells ＋ stromal cells ＋ aVLA-4 ＋ Ara-C group （antibody ＋ drug group）. Cell proliferation or inhibition rate was detected by CCK-8 method, the HL-60 cells apoptosis was detected by flow cytometry. The expression of anti-apoptotic gene bcl-2 in HL-60 cells was determined by Western blot. Results After 24 h and 48 h, treatment, the number of the stromal cells group HL-60 cells were higher than that of the control group with significant difference cultured [（7.2±0.3）×l0^5/ml vs （5.3_＋0.4）×l0^5/ml, （8.4＋0.2）×l0^5/ml vs （6.8±0.3）×l0^5/ml, P 〈 0.001], while the HL-60 cell proliferation inhibition rate [（24.3±2.1） %, （37.0±2.6） %, （65.6±3.8）%] and apoptosis rate [（5.7±0.6） %, （8.0±0.5） %, （10.4±0.9） %, （16.5±0.7） %] of antibody group, drug group, antibody ＋ drug group were higher than the control group with a difference of statistically significant （P 〈 0.05）, and the increase of antibody ＋ drug group was most obvious. With the decreasing of the bcl-2 protein expression, which was most the decrease of antibody ＋ drug group was most obvious. Conclusion Bone marrow stromal cells can stimulate the proliferation of leukemia cells, aVLA-4 interference the interaction between stromal cells and leukemia cells can enhance the chemosensitivity of leukemia cells to Ara-C.