摘要
目的探讨苦参素注射液(matrine injection,MI)联合顺铂(cisplatin,DDP)对人肝癌SMMC-7721细胞增殖抑制作用及其机制。方法采用四甲基偶氮唑盐(MTT)法检测细胞抑制率,流式细胞仪(FCM)检测细胞周期分布,半定量RT-PCR法检测c-myc mRNA表达,二步法免疫组化检测c-Myc、P27和CyclinD1蛋白表达。结果 MI、DDP单独应用均对人肝癌SMMC-7721细胞有抑制作用(P<0.01或P<0.05),联合用药具有协同效应。与单药组相比,联合用药组的细胞抑制率显著上升(P<0.05或P<0.01),G0/G1期细胞比例显著升高(P<0.05),S期细胞比例明显减少(P<0.05),P27表达显著上升(P<0.05),c-myc mRNA和蛋白、CyclinD1表达显著降低(P<0.05)。结论 MI联合DDP具有明显的协同抑制肝癌SMMC-7721细胞增殖的作用,使细胞周期阻滞于G0/G1期,其机制可能与上调P27、下调c-myc基因和CyclinD1表达有关。
【Objective】 To investigate the proliferation inhibitory effects and its mechanism of matrine injection combined with cisplatin on human hepatocarcinoma cell line.【Method】 The inhibitory rat was evaluated by MTT assay.The cell cycle distribution was examined by flow cytometry.The expression of c-myc mRNA was determined by semi-quantitive RT-PCR,and the expression of c-Myc,P27and CyclinD1 proteins was detected by two-step immunhistochemical staining.【Results】 Matrine injection and cisplatin used alone could inhibit the proliferation of SMMC27721 cells（P 0.05 or P 0.01）,and there was the synergistic effect after treated with combined agents.Compared with single drug group,the inhibitory rate was increased significantly（P 0.05 or P 0.01）,the number of cells in G0/G1 phase was increased significantly（P 0.05）,while those in S phase was decreased significantly（P 0.05）,the expression of P27 protein was increased significantly（P 0.05）,the expression of c-myc mRNA and protein,CyclinD1 protein was decreased markedly（P 0.05）.【Conclusion】 Matrine injection combined with cisplatin can obviously inhibit the proliferation of SMMC27721 cells and displays the synergistic effect.The cell cycle is arrested at G0/G1 phase.Its mechanism may be involved in up-regulating expression of P27 and down-regulating expression of c-myc gene and CyclinD1.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2012年第31期36-40,共5页
China Journal of Modern Medicine
基金
广西自然科学基金(No:0542119)
广西高校优秀人才资助项目(No:桂教人20077029)
广西卫生厅重点课题(No:重200887)
广西教育厅课题(No:桂教科研200510197)
