消癌平辅助丝裂霉素对肺癌细胞化疗增效的研究

Effects of Xiaoaiping injection combined with mitomycin on lung cancer cells in vitro

目的通过消癌平联合丝裂霉素对肺癌A549细胞增殖,细胞周期、凋亡及相关基因表达的影响,探讨化疗增效机制。方法 MTT检测消癌平(2、8、21μl/ml)联合丝裂霉素(2μg/nl)对肺癌A549细胞生长抑制(联合用药A、B、C组),并设立对照组、丝裂霉素组和消癌平组;流式细胞术检测细胞周期及凋亡;RT-PCR检测P53、Cyclin D1基因表达。结果 MTT检测显示在24、48、72 h联合用药A、B、C组OD(光密度)均比丝裂霉素低(P<0.05);流式细胞术检测表明消癌平、丝裂霉素与对照组相比,肺癌细胞均阻滞于G0/G1期,差异有统计学意义(P<0.05)。联合用药组G0/G1期细胞比率、细胞凋亡率与丝裂霉素组比较,差异有统计学意义(P<0.05)。消癌平组、丝裂霉素组与对照组比较,均能提高P53 mRNA表达(P<0.05),联合治疗组P53 mRNA表达高于丝裂霉素组(P<0.05)。消癌平组、丝裂霉素组与对照组比较均能降低Cyclin D1基因的表达(P<0.05)。与丝裂霉素组比较,联合用药可显著减低Cyclin D1基因的表达(P<0.05)。结论消癌平辅助丝裂霉素对肺癌细胞化疗增效可能与上调P53 mRNA表达,增强细胞凋亡作用,以及下调Cyclin D1 mRNA表达,阻滞细胞周期于G0/G1期有关。

Objective To observe the effects of Xiaoaiping injection combined with mitomycin on cell proliferation,cell cycle,cell apoptosis and expressions of related genes( Cyclin D1,P53 mRNA) of lung cancer-A549 cells in vitro,and to explore potential action mechanism. Methods The inhibitory effects of Xiaoaiping injection( 2,8,21μl / ml) combined with mitomycin( 2μg / nl) were detected by MTT( combination treatment group A,B,C),moreover,control group,Xiaoaiping group and mitomycin group were established. The cell cycle and cell apoptosis of lung cancer-A549 cells in vitro were detected by cytometry,and the expression levels of Cyclin D1 and P53 mRNA were measured by RT-PCR. Results The results by MTT showed that the optical density value( OD) on 24 h,48h,72 h in combination treatment group A,B,C was significantly higher than that in mitomycin group( P < 0. 05). The results by cytometry showed that as compared with control group,the lung cancer cells were blocked at G0 / G1 phase in Xiaoaiping group and mitomycin group( P < 0. 05). There were significant differences in the cell ratio at G0 / G1 phase and cell apoptosis rate between combination treatment groups and mitomycin group( P < 0. 05). The expression levels of P53 mRNA in Xiaoaiping group and mitomycin group were significantly increased,as compared with those in control group( P < 0. 05). Moreover the expression levels of P53 mRNA in combination treatment groups were significantly higher than those in mitomycin group( P < 0. 05). The expression levels of Cyclin D1 in Xiaoaiping group and mitomycin group were significantly decreased,as compared with those in control group( P < 0. 05). Furthermore the expression levels of Cyclin D1 in combination treatment groups were significantly decreased,as compared with those in mitomycin group( P < 0. 05). Conclusion Xiaoaiping injection combined with mitomycin can enhance the efficiency of chemotherapy on lung cancer-A549 cells in vitro,and the action mechanism may be correlated to up- regulating the expression levels of P53 mRNA,increasing cell apoptosis and down- regulating the expressions of Cyclin D1 mRNA to block cancer cells at G0 / G1 phase.