低相对分子质量肝素增强顺铂诱导肝癌细胞的凋亡作用

目的：探讨低相对分子质量肝素(low molecular weight heparin，LMWH)对顺铂(cisplatin，DDP)诱导肝癌细胞凋亡作用的影响及其作用机制。方法：实验分为对照组、LMWH组、DDP组、LWMH与DDP联合应用组。应用MMT法检测药物对肝癌SMMC-7721细胞增殖的影响；吖啶橙/溴化乙锭双荧光法和流式细胞术检测各组细胞凋亡情况；实时荧光定量PCR和Western印迹检测细胞凋亡相关蛋白Fas，Bcl-2和Bax的表达。结果：MTT检测结果显示：与对照组相比，DDP组及LWMH与DDP联合应用组的SMMC-7721细胞生长均有显著的抑制(均P<0.05)；LMWH组的Fas，Bcl-2和Bax mRNA及蛋白均无明显的变化(均P>0.05)；DDP组Fas表达量明显增加(P<0.05)，而Bcl-2轻度降低(P<0.05)，Bax 无明显变化(P>0.05)。LWMH与DDP联合应用组与对照组及DDP组相比，Bcl-2表达明显降低(均P<0.05)，Bax表达明显增高(均P<0.05)；Fas表达量与对照组相比明显增加(P<0.05)，但与DDP组相比增加不明显(P>0.05)。结论：LMWH具有增强DDP诱导肝癌SMMC-7721细胞凋亡的作用，其机制可能与LMWH增强DDP诱导的细胞凋亡中线粒体通路和促使细胞凋亡增加有关。

Potentiation of cisplatin induced apoptosis by low molecular weight heparin in human hepatocellular carcinoma cell

Objective: To explore the effects of low molecular weight heparin (LMWH) on cisplatin (DDP)- induced apoptosis in humanhepatocellularcarcinomacell and the underlying mechanisms.Methods: Hepatocellular carcinoma SMMC-7721 cells were divided into a control group, a LMWH group, a DDP group and a LMWH plus DDP group. The effect of the drugs on proliferation of hepatocellular carcinoma SMMC-7721 cells were evaluated by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide (MTT) assay, and the apoptosis were detected by acridine orange/ethidium bromide (AO/EB) double fluorescence method and flow cytometry method. The expression levels of apoptosis-related protein Fas, Bcl-2 and Bax were detected by quantitative real-time PCR and Western blot.Results: Compared with the control group, the proliferation of SMMC-7721 cells was inhibited in the DDP group and the LMWH plus DDP group (both P<0.05). The mRNA and protein levels of Fas, Bcl-2, Bax in the SMMC-7721 cells were not obviously changed in the LMWH group (all P>0.05). The Fas level was increased obviously (P<0.05), while the Bcl-2 level moderately reduced (P<0.05), Bax were not obviously changed in the DDP group (P>0.05). Compared with the control group and the DDP group, the Bcl-2 level was reduced significantly in the LMWH plus DDP group (both P<0.05), while the level of Bax was increased obviously (both P<0.05). Compared with control group, the Fas level was increased significantly in the LMWH plus DDP group (P<0.05), but the increase was not significant compared with the DDP group (P>0.05).Conclusion: LMWH can enhance the cisplatin-induced apoptosis in SMMC-7721 cells, which might be related to activation of mitochondrial pathway.