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

作　　者：吴宣宣 张俊丽 商玲 燕善军[1] 武文娟[3] WU Xuan-xuan;ZHANG Jun-li;SHANG Ling;YAN Shan-jun;WU Wen-juan(Department of Gastroenterology,The First Affiliated Hospital of Bengbu Medical College,Bengbu Anhui 230004;Experimental Center of Clinical Laboratory Diagnostics,Bengbu Medical College,Bengbu Anhui 233030,China;Department of Biochemistry and Molecular Biology,Bengbu Medical College,Bengbu Anhui 233030,China)

机构地区：[1]蚌埠医学院第一附属医院消化科,安徽蚌埠233004 [2]蚌埠医学院临床检验诊断学实验中心,安徽蚌埠233030 [3]蚌埠医学院生物化学与分子生物学教研室,安徽蚌埠233030

出　　处：《蚌埠医学院学报》2023年第4期426-431,共6页Journal of Bengbu Medical College

基　　金：安徽省教育厅自然科学研究项目(KJ2017A234)。

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。Objective:To 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.Methods:The 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.Results:Compared 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 LC3Ⅱsignificantly 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).Conclusions:The LMWH combined with DDP can significantly increase the autophagy of BEL-7404 cells,which may be realized through activating AMPK-ULK1-Beclin1 signaling pathway.

关 键 词：肝肿瘤 低分子质量肝素 顺铂 自噬 AMPK-ULK1-Beclin1 

分 类 号：R735.7[医药卫生—肿瘤]