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目的探讨吉西他滨对体外培养的人骨肉瘤细胞增殖的影响及其可能的作用机制。方法体外传代培养人骨肉瘤MG-63细胞,实验时取对数生长期的细胞。将不同浓度(0.025、0.050、0.100、0.200、0.400 mg/L)的吉西他滨分别作用于MG-63细胞24 h,用四甲基偶氮唑盐(MTT)法检测其对MG-63细胞的生长抑制率。用实时定量聚合酶链反应(PCR)检测吉西他滨作用24 h后人骨肉瘤MG-63细胞肿瘤转移相关基因1(MTA1)mRNA的相对表达量。结果 0.025、0.050、0.100、0.200、0.400 mg/L吉西他滨作用24 h后,MG-63细胞抑制率分别为(3.3±1.2)%、(3.7±1.0)%、(6.1±0.1)%、(8.0±1.9)%和(11.5±1.4)%,差异有统计学意义(F=30.568,P=0.0169);其中0.025、0.050 mg/L组MG-63抑制率均低于其他3组,0.100、0.200 mg/L组亦低于0.400 mg/L组(P<0.05)。0.100、0.200、0.400 mg/L吉西他滨作用于人骨肉瘤MG-63细胞24 h后,MTA1 mRNA相对表达量〔分别为(0.86±0.16)、(0.68±0.12)、(0.46±0.08)〕均低于空白对照组(1.01±0.15),且3个剂量吉西他滨组MTA1mRNA相对表达量依次降低,差异有统计学意义(P<0.01)。结论吉西他滨能抑制人骨肉瘤MG-63细胞增殖,且有剂量依赖性;其作用机制可能是通过下调MTA1 mRNA的表达,从而抑制MG-63细胞转移,发挥抗骨肉瘤作用。
Objective To investigate the effects of gemcitabine on the proliferation of human osteosarcoma cells cultured in vitro and its possible mechanism. Methods Human osteosarcoma MG-63 cells were subcultured in vitro, and the logarithmic growth phase cells were used in the experiment. Gemcitabine at different concentrations (0.025,0.050,0.100,0.200,0.400 mg / L) were applied to MG-63 cells for 24 h, respectively. The growth of MG-63 cells was detected by MTT assay Inhibition rate. The relative expression of MTA1 mRNA in human osteosarcoma MG-63 cells was detected by real-time polymerase chain reaction (PCR) 24 hours after gemcitabine treatment. Results The inhibitory rates of MG-63 cells after (0.025,0.050,0.100,0.200,0.400 mg / L) gemcitabine treatment for 24 h were (3.3 ± 1.2)%, (3.7 ± 1.0)%, (6.1 ± 0.1)%, (F = 30.568, P = 0.0169). The inhibitory rates of MG-63 in 0.025 and 0.050 mg / L groups were all lower than those in the other three groups, 0.100 and 0.200 mg / L group was also lower than 0.400 mg / L group (P <0.05). The relative expression of MTA1 mRNA (0.86 ± 0.16, 0.68 ± 0.12, (0.46 ± 0.08), respectively] was lower than 0.100,0.200,0.400 mg / L gemcitabine for 24 h in human osteosarcoma MG-63 cells The blank control group (1.01 ± 0.15), and the relative expression of MTA1 mRNA in three doses of gemcitabine group in turn decreased, the difference was statistically significant (P <0.01). Conclusion Gemcitabine inhibits the proliferation of human osteosarcoma MG-63 cells in a dose-dependent manner. The possible mechanism is that gemcitabine inhibits the metastasis of MG-63 cells by down-regulating the expression of MTA1 mRNA and playing the role of anti-osteosarcoma.