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目的研究不同种类的非甾体类抗炎药(NSAIDs)对人骨肉瘤细胞株增殖和凋亡的影响。方法采用有限稀释法,从人骨肉瘤细胞系MG-63中分离培养出细胞株,扩增培养后分为4个干预组:Cox-1特异性抑制剂组(小剂量阿司匹林,采用的浓度梯度分别为:0μg/ml、5μg/ml、10μg/ml、15μ/ml、20μ/ml、25μg/ml)、Cox非选择性抑制剂组(双氯芬酸钠注射液,采用的浓度梯度分别为:0mg/L、0.3mg/L、0.6mg/L、0.9mg/L、1.2mg/L、1.5mg/L)、Cox-2选择性抑制剂组(美洛昔康注射液,采用的浓度梯度分别为:0μg/ml、0.4μg/ml、0.8μg/ml、1.2μg/ml、1.6μg/ml、2.0μg/m1)、Cox-2特异性抑制剂组(注射用帕瑞昔布,采用的浓度梯度分别为:0mg/L、0.3mg/L、0.6mg/L、0.9mg/L、1.2mg/L、1.5mg/L),设培养液中不加药物的作为对照组,每组中的不同浓度均设6个重复孔,药物对细胞株干预72h后,采用四唑盐比色法(MTT)检测细胞增殖抑制率,流式细胞仪测定细胞凋亡率。结果不同浓度的小剂量阿司匹林、双氯芬酸钠、美洛昔康、帕瑞昔布作用于骨肉瘤细胞株后均能不同程度地抑制细胞株的生长,各处理组与对照组之间具有显著性差异(P<0.05),在常见的血药浓度范围内,抑制率与浓度之间成正相关(P<0.05);不同浓度的小剂量阿司匹林、双氯芬酸钠、美洛昔康、帕瑞昔布也均能不同程度地促使细胞株凋亡,实验组的凋亡率明显高于对照组,其差异具有统计学意义(P<0.05),在常见血药浓度的范围内,浓度越高,凋亡率就越高(P<0.05)。结论 4种不同种类的NSAIDs均能抑制骨肉瘤细胞的增殖和促使其凋亡,这种作用具有量效依赖关系。
Objective To study the effects of different kinds of non-steroidal anti-inflammatory drugs (NSAIDs) on the proliferation and apoptosis of human osteosarcoma cell lines. Methods The cell strains were isolated and cultured from human osteosarcoma cell line MG-63 using limited dilution method. After expansion, the cells were divided into four groups: Cox-1 specific inhibitor group (low-dose aspirin, concentration gradients (Diclofenac sodium injection: 0 mg / L, 0 μg / ml, 5μg / ml, 10μg / ml, 15μ / ml, 20μ / ml and 25μg / ml respectively) , 0.3mg / L, 0.6mg / L, 0.9mg / L, 1.2mg / L, 1.5mg / L) and Cox-2 selective inhibitor (meloxicam injection, Cox-2-specific inhibitor group (parecoxib for injection, concentration gradient used Respectively, 0mg / L, 0.3mg / L, 0.6mg / L, 0.9mg / L, 1.2mg / L, 1.5mg / L), set the culture medium without drugs as a control group, Six replicate wells were set in each group. After 72 h of drug intervention, cell proliferation inhibition rate was detected by tetrazolium salt (MTT) assay and apoptosis rate was determined by flow cytometry. Results The low dose aspirin, diclofenac sodium, meloxicam, and parecoxib could inhibit the growth of osteosarcoma cell lines to a certain extent after treatment with different concentrations of aspirin, and there was a significant difference between the treatment groups and the control group (P <0.05). In the range of common plasma concentration, there was a positive correlation between the inhibition rate and the concentration (P <0.05). Low concentrations of aspirin, diclofenac sodium, meloxicam and parecoxib The apoptosis rate of experimental group was significantly higher than that of control group (P <0.05). In the range of common plasma concentration, the higher the concentration, the higher the apoptosis rate The higher (P <0.05). Conclusion All four different kinds of NSAIDs can inhibit osteosarcoma cell proliferation and induce apoptosis, and this effect has dose-dependent relationship.