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    吴宣宣, 张俊丽, 商玲, 燕善军, 武文娟. 低分子质量肝素联合顺铂对肝癌细胞自噬的影响[J]. 蚌埠医科大学学报, 2023, 48(4): 426-431. DOI: 10.13898/j.cnki.issn.1000-2200.2023.04.002
    引用本文: 吴宣宣, 张俊丽, 商玲, 燕善军, 武文娟. 低分子质量肝素联合顺铂对肝癌细胞自噬的影响[J]. 蚌埠医科大学学报, 2023, 48(4): 426-431. DOI: 10.13898/j.cnki.issn.1000-2200.2023.04.002
    shu
    WU Xuan-xuan, ZHANG Jun-li, SHANG Ling, YAN Shan-jun, WU Wen-juan. Effect of the low molecular weight heparin combined with cisplatin on the autophagy of liver cancer cells[J]. Journal of Bengbu Medical University, 2023, 48(4): 426-431. DOI: 10.13898/j.cnki.issn.1000-2200.2023.04.002
    Citation: WU Xuan-xuan, ZHANG Jun-li, SHANG Ling, YAN Shan-jun, WU Wen-juan. Effect of the low molecular weight heparin combined with cisplatin on the autophagy of liver cancer cells[J]. Journal of Bengbu Medical University, 2023, 48(4): 426-431. DOI: 10.13898/j.cnki.issn.1000-2200.2023.04.002
    shu

    低分子质量肝素联合顺铂对肝癌细胞自噬的影响

    Effect of the low molecular weight heparin combined with cisplatin on the autophagy of liver cancer cells

      摘要
    • 摘要:
      目的探讨低分子质量肝素(low molecular weight heparin, LMWH)联合顺铂(cisplatin, DDP)对BEL-7404人肝癌细胞自噬的影响和机制。
      方法用不同质量浓度的LMWH、DDP单独或联合处理肝癌BEL-7404细胞48 h, 并分为4组: 对照组、LMWH组、DDP组及L+D组。CCK-8法测其对细胞增殖能力的影响; 单丹磺酰尸胺荧光染色法观察自噬水平; RT-PCR检测自噬蛋白mRNA的表达; Western blotting检测自噬蛋白及通路蛋白的表达。
      结果L+D组的细胞增殖能力较其他组明显下降(P < 0.01), 绿色荧光自噬体显著增多、自噬相关蛋白LC3Ⅱ的mRNA和蛋白表达均明显增高(P < 0.01), p62明显降低(P < 0.01), P-AMPK/AMPK、P-ULK1/ULK1以及Beclin1的蛋白表达明显升高(P < 0.05~P < 0.01)。
      结论LMWH与DDP联合作用显著提高了BEL-7404细胞自噬, 可能是通过激活AMPK-ULK1-Beclin1信号通路而实现。

       

      Abstract:
      ObjectiveTo investigate the effects and mechanism of low molecular weight heparin(LMWH) combined with cisplatin(DDP) on the autophagy of human hepatocellular carcinoma(HCC) BEL-7404 cells.
      MethodsThe BEL-7404 liver cancer cells were treated with different concentrations of LMWH and DDP alone or in combination for 48 h, and divided into the control group, LMWH group, DDP group and L+D group.CCK-8 method was used to measure the effects on cell proliferation ability.Dansylcadaverine fluorescence staining was used to observe the autophagy levels.The expression of autophagy-related protein mRNA was detected by RT-PCR.Western blotting was used to detect the expression of autophagy-related proteins and pathway proteins.
      ResultsCompared with other groups, the cell proliferation ability significantly decreased(P < 0.05), the green fluorescence autophagy body significantly increased, the mRNA and protein expression levels of autophagy-related protein LC3Ⅱ significantly increased(P < 0.01), the p62 significantly decreased(P < 0.01), and the protein expressions of P-AMPK/AMPK, P-ULK1/ULK1 and Beclin1 significantly increased in the L+D group(P < 0.05 to P < 0.01).
      ConclusionsThe LMWH combined with DDP can significantly increase the autophagy of BEL-7404 cells, which may be realized through activating AMPK-ULK1-Beclin1 signaling pathway.

       

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