甘草内生菌联合顺铂对A549细胞的协同抑制作用研究

Synergistic inhibitory effect of licorice endophyte combined with cisplatin on A549 cells

目的探讨甘草内生菌JTYB34联合顺铂对人肺癌A549细胞协同抑制作用及其可能机制。方法将肺癌A549细胞分为对照组(正常培养)、顺铂组(4μg·mL^(-1)顺铂)、JTYB34组(160μg·mL^(-1)JTYB34)及联合组(4μg·mL^(-1)顺铂联合160μg·mL^(-1)JTYB34)。用噻唑蓝法检测A549细胞存活率,用流式细胞术检测A549细胞凋亡率,用蛋白质印迹法检测A549细胞凋亡关键蛋白的表达水平。结果对照组、顺铂组、JTYB34组和联合组的细胞存活率分别为(100.00±3.53)%、(86.95±1.74)%、(64.98±1.36)%和(37.06±1.18)%,细胞凋亡率分别为(4.89±0.26)%、(10.15±0.26)%、(10.76±0.07)%和(20.43±0.16)%,B细胞淋巴瘤/白血病-2(Bcl-2)蛋白相对表达水平分别为0.99±0.03、0.85±0.01、0.71±0.01和0.59±0.02,Bcl-2相关X蛋白相对表达水平分别为0.72±0.01、0.82±0.01、1.27±0.02和1.33±0.02,细胞色素C蛋白相对表达水平分别为0.47±0.00、0.83±0.01、0.89±0.01和1.08±0.03。顺铂组和JTYB34组的上述指标与对照组比较,联合组的上述指标与各单药组比较,差异均有统计学意义(P<0.05,P<0.01)。结论顺铂和JTYB34能够抑制肺癌细胞增殖,促进肺癌细胞凋亡,两药联合使用具有协同增效作用。

Objective To investigate the synergistic inhibitory effect of licorice endophyte JTYB34 combined with cisplatin on human lung cancer A549 cells and its possible mechanism.Methods Lung cancer A549cells were divided into control group(normal culture),cisplatin group(4μg·m L^(-1)cisplatin),JTYB34 group(160μg·m L^(-1)JTYB34)and combination group(4μg·m L^(-1)cisplatin combined with 160μg·m L^(-1)JTYB34).The survival rate of A549 cells was detected by methyl thiazolyl tetrazo-lium method,the apoptosis rate was detected by flow cytometry,and the expression level of key proteins in apoptosis of A549cells was detected by Western blot.Results The cell survival rates of control group,cisplatin group,JTYB34 group and combination group were(100.00±3.53)%,(86.95±1.74)%,(64.98±1.36)%and(37.06±1.18)%,respectively;the apoptosis rates were(4.89±0.26)%,(10.15±0.26)%,(10.76±0.07)%and(20.43±0.16)%,respectively;the relative expression levels of B cell lymphoma/leukemia-2(Bcl-2)protein were 0.99±0.03,0.85±0.01 and 0.71±0.01 and 0.59±0.02,respectively;the relative expression levels of Bcl-2 associated X protein were 0.72±0.01,0.82±0.01,1.27±0.02 and 1.33±0.02,respectively;the relative expression levels of cytochrome C protein were 0.47±0.00,0.83±0.01,0.89±0.01 and 1.08±0.03,respectively.There were statistically significant differences in the above indexes between cisplatin group,JTYB34 group and control group,and between combination group and monotherapy groups(P<0.05,P<0.01).Conclusion Cisplatin and JTYB34 can inhibit the proliferation of lung cancer cells and promote the apoptosis of lung cancer cells.The combination of the two drugs has a synergistic effect.