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目的:探讨人急性早幼粒白血病细胞系(HL-60)和其耐药株抗阿霉素人急性早幼粒白血病细胞系(HL-60/ADM)的超弱发光与耐药性的关系。方法:应用MTT比色法、流式细胞术、细胞生长曲线和免疫组化染色法,检测HL-60细胞、HL-60/ADM细胞对阿霉素(ADM)的敏感性、细胞周期变化及P-糖蛋白(P-gp170)的表达。用IFFM-D型流动式化学发光仪检测HL-60细胞及HL-60/ADM细胞的超弱发光强度。结果:HL-60/ADM细胞对ADM的敏感性明显低于亲本HL-60细胞;HL-60/ADM细胞中G0/G1期的细胞为44·80%±1·97%,G2/M期的细胞为9·90%±0·27%,较其亲本细胞增多,S期的细胞为45·30%±1·93%,较其亲本细胞减少(P<0·05);HL-60/ADM细胞倍增时间是HL-60细胞的1·8倍,P-gp170的表达呈阳性,HL-60细胞呈阴性。在1×10-4mol/L鲁米诺及3mL/L双氧水条件下,密度分别为8×107/L、1×108/L、1·26×108/L及2·73×108/L的HL-60/ADM细胞超弱发光强度低于HL-60细胞(P<0·05)。结论:HL-60/ADM细胞对ADM的敏感性降低,细胞生长延缓,P-gp170表达呈阳性等均提示HL-60/ADM细胞出现了耐药性;且随着HL-60/ADM细胞耐药性的产生其超弱发光强度较其亲本细胞HL-60降低,提示细胞超弱发光强度明显降低可作为细胞耐药性产生的一项指标。
Objective: To investigate the relationship between ultra-weak chemoresistance and human acute promyelocytic leukemia cell line (HL-60) and its multidrug-resistant strains against adriamycin-induced human acute promyelocytic leukemia cell line HL-60 / ADM . Methods: MTT assay, flow cytometry, cell growth curve and immunohistochemical staining were used to detect the sensitivity of adriamycin (ADM) and cell cycle to HL-60 cells and HL-60 / ADM cells P-glycoprotein (P-gp170) expression. The ultra-weak luminescence intensity of HL-60 cells and HL-60 / ADM cells was detected by IFFM-D flow cytometer. RESULTS: The sensitivity of HL-60 / ADM cells to ADM was significantly lower than that of the parental HL-60 cells. The cells in G0 / G1 phase of HL-60 / ADM cells were 44.8% ± 1.97% (P <0.05). The cell number of HL-60 cells was 9.90% ± 0.27%, which was 45.30% ± 1.93% in S phase compared with their parental cells / ADM cell doubling time is 1.8 times HL-60 cells, P-gp170 expression was positive, HL-60 cells were negative. Under the conditions of 1 × 10-4mol / L luminol and 3mL / L H2O2, the densities were 8 × 107 / L, 1 × 108 / L, 1.26 × 108 / L and 2.73 × 108 / L respectively The ultra-weak luminescence intensity of HL-60 / ADM cells was lower than HL-60 cells (P <0.05). CONCLUSION: The sensitivity of HL-60 / ADM cells to ADM is decreased, the cell growth is delayed and the expression of P-gp170 is positive, which indicates the drug resistance of HL-60 / ADM cells. Its ultra-weak luminescence intensity is lower than that of its parental cell HL-60, which indicates that the ultra-weak luminescence intensity of cells is significantly reduced as an indicator of cell resistance.