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中国精品科技期刊2020
陈永芳,李艳珂,张淑静. 茯苓多糖通过LncRNA HCG11/miR-539-3p调控肝癌细胞自噬、化疗耐药的机制[J]. 华体会体育,2024,45(22):322−330. doi: 10.13386/j.issn1002-0306.2023110291.
引用本文: 陈永芳,李艳珂,张淑静. 茯苓多糖通过LncRNA HCG11/miR-539-3p调控肝癌细胞自噬、化疗耐药的机制[J]. 华体会体育,2024,45(22):322−330. doi: 10.13386/j.issn1002-0306.2023110291.
CHEN Yongfang, LI Yanke, ZHANG Shujing. Mechanism of Poria cocos Polysaccharide Regulating Autophagy and Chemotherapy Resistance of Hepatocellular Carcinoma Cells through LncRNAHCG11/miR-539-3p 539-3p[J]. Science and Technology of Food Industry, 2024, 45(22): 322−330. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023110291.
Citation: CHEN Yongfang, LI Yanke, ZHANG Shujing. Mechanism of Poria cocos Polysaccharide Regulating Autophagy and Chemotherapy Resistance of Hepatocellular Carcinoma Cells through LncRNAHCG11/miR-539-3p 539-3p[J]. Science and Technology of Food Industry, 2024, 45(22): 322−330. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023110291.

茯苓多糖通过LncRNA HCG11/miR-539-3p调控肝癌细胞自噬、化疗耐药的机制

Mechanism of Poria cocos Polysaccharide Regulating Autophagy and Chemotherapy Resistance of Hepatocellular Carcinoma Cells through LncRNAHCG11/miR-539-3p 539-3p

  • 摘要: 目的:探讨茯苓多糖(Poria cocos polysaccharide,PCP)对肝癌细胞增殖、自噬和化疗耐药的影响及机制。方法:对数期HepG-2/DDP细胞随机分组:对照组、PCP组(7.5、15、30 µg/mL PCP)、shNC 组、HCG11 siRNA组(转染50 nmol/L HCG1干扰序列)、pcDNA组(转染2 µg pcDNA)、pcDNA3.1-HCG11组(转染2 µg pcDNA3.1-HCG11)、PCP组(30 µg/mL)和PCP+pcDNA3.1-HCG11组(转染2 µg pcDNA3.1-HCG11+30 µg/mL PCP)。MTT检测肝癌细胞耐药情况和抑制率,Western blot检测Beclin-1、LC3 I和LC3 II水平,实时荧光定量PCR(Real time quantitative PCR,qRT-PCR)检测细胞株中LncRNA HCG11和miR-539-3p的水平,双荧光素酶报告实验检测LncRNA HCG11和miR-539-3p的靶向关系,裸鼠成瘤实验验证体内瘤体的瘤重、瘤体积及自噬相关蛋白水平。结果:4~128 µg/mL PCP抑制HepG-2/DDP细胞增殖,7.5、15、30 µg/mL的PCP能显著降低HepG-2/DDP细胞中MRP1、P-gp、Beclin-1、LC3II的水平,下调HCG11水平,增加miR-539-3p的表达水平(P<0.01);与对照组比较,HCG11 siRNA 组HepG-2/DDP细胞HCG11相对表达水平、IC50(顺铂)、Beclin-1、LC3 II/ LC3 I比值显著降低,miR-539-3p水平和抑制率(顺铂)显著增加(P<0.01);与PCP组比较,PCP+pcDNA3.1-HCG11处理组HCG11相对表达水平显著升高,Belin-1、LC3 II/LC3 I比值显著增加(P<0.01);在不同浓度的顺铂处理下,与PCP组比较,PCP+pcDNA3.1-HCG11 组抑制率显著降低(P<0.01);与PCP组比较,PCP+HCG11 shRNA 组肿瘤体积和肿瘤重量显著降低,Belin-1、LC3 II/LC3 I水平显著下调(P<0.05)。结论:PCP能通过LncRNA HCG11/miR-539-3p调控耐药和自噬,并影响肝癌细胞的增殖。

     

    Abstract: Objective: To explore the effects and mechanism of Poria cocos polysaccharide (PCP) on proliferation, autophagy and chemotherapy resistance of hepatocellular carcinoma cells. Methods: HepG-2/DDP cells in logarithmic phase were randomly divided into control group, PCP group (7.5, 15, 30 μg/mL PCP), shNC group, HCG11 siRNA group (transfection of 50 nmol/L HCG1 interference sequence), pcDNA group (transfection of 2 μg pcDNA), pcDNA3.1-HCG11 group (transfection of 2 μg pcDNA3.1-HCG11), PCP group (30 μg/mL PCP) and PCP+pcDNA3.1-HCG11 group (transfection of 2 μg pcDNA3.1-HCG11+30 μg/mL PCP). MTT assay was used to detect the drug resistance and inhibition rate of hepatocellular carcinoma cells, western blot was used to detect the levels of Beclin-1, LC3, and quantitative real time polymerase chain reaction (qRT-PCR) was used to detect the levels of LncRNA HCG11 and miR-539-3p in the cell lines. Double luciferase report experiment was used to detect the targeting relationship between LncRNA HCG11 and miR-539-3p, and nude mice tumorigenesis experiment was used to verify the influence of tumor weight, tumor volume and autophagy-related protein level in nude mice. Results: 4~128 µg/mL PCP had a certain inhibitory effect on HepG-2/DDP cells. PCP of 7.5, 15, 30 μg/mL could significantly decrease the levels of MRP1, P-gp, Beclin-1 and LC3II, decrease the level of HCG11 and increase the expression of miR-539-3p in HepG-2/DDP cells (P<0.01). Compared with the control group, the relative expression level of HCG11, IC50 (cisplatin), Beclin-1 and LC3II/LC3I in HCG11siRNA group decreased significantly, while the level of miR-539-3p and inhibition rate (cisplatin) increased significantly (P<0.01). Compared with PCP group, the relative expression level of HCG11 in PCP+pcDNA3.1-HCG11 group increased significantly, while the ratio of Belin-1 and LC3 II/LC3 I increased significantly (P<0.01). Compared with PCP group, the inhibition rate of PCP+pcDNA3.1+HCG11 group was significantly decreased under different concentrations of cisplatin (P<0.01). Compared with PCP group, tumor volume and tumor weight in PCP+HCG11shRNA group were significantly decreased, and the levels of Belin-1 and LC3II/LC3I were significantly decreased (P<0.05). Conclusion: PCP can affect the proliferation of hepatocellular carcinoma cells by regulating drug resistance and autophagy through LncRNA HCG11/miR-539-3p.

     

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