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

作　　者：陈永芳[1] 李艳珂 张淑静[1] CHEN Yongfang;LI Yanke;ZHANG Shujing(Luohe Medical College,Luohe 462002,China)

机构地区：[1]漯河医学高等专科学校,河南漯河462002

出　　处：《食品工业科技》2024年第22期322-330,共9页Science and Technology of Food Industry

基　　金：河南省高等学校重点科研项目(18A31001)。

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、30μg/mL PCP)、shNC组、HCG11siRNA组(转染50 nmol/L HCG1干扰序列)、pcDNA组(转染2μg pcDNA)、pcDNA3.1-HCG11组(转染2μg pcDNA3.1-HCG11)、PCP组(30μg/mL)和PCP+pcDNA3.1-HCG11组(转染2μg pcDNA3.1-HCG11+30μg/mL PCP)。MTT检测肝癌细胞耐药情况和抑制率,Western blot检测Beclin-1、LC3Ⅰ和LC3Ⅱ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:4~128μg/mL PCP抑制HepG-2/DDP细胞增殖,7.5、15、30μg/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、IC_(50)(顺铂)、Beclin-1、LC3Ⅱ/LC3Ⅰ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、LC3Ⅱ/LC3Ⅰ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、LC3Ⅱ/LC3Ⅰ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。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(7.5,15,30μg/mL PCP),shNC group,HCG11 siRNA group(transfection of 50 nmol/L HCG1 interference sequence),pcDNA group(transfection of 2μg pcDNA),pcDNA3.1-HCG11 group(transfection of 2μg pcDNA3.1-HCG11),PCP group(30μg/mL PCP)and PCP+pcDNA3.1-HCG11 group(transfection of 2μg pcDNA3.1-HCG11+30μg/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:4~128µg/mL PCP had a certain inhibitory effect on HepG-2/DDP cells.PCP of 7.5,15,30μg/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,IC_(50)(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 tumo

关 键 词：茯苓多糖 LncRNA HCG11 miR-539-3p 化疗耐药性 自噬 

分 类 号：Q949.9[生物学—植物学]