首页|白花蛇舌草提取物调控微小RNA-340对肝癌细胞增殖、迁移及侵袭的影响

白花蛇舌草提取物调控微小RNA-340对肝癌细胞增殖、迁移及侵袭的影响

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目的 观察白花蛇舌草提取物(HDE)对肝癌细胞增殖、迁移、侵袭的作用及机制.方法 将肝癌细胞HCC-7721、LM3分为对照组(NC组)、HDE低剂量组(HDE-L组)和HDE高剂量组(HDE-H组),并予以相应的药物干预48 h,分别以CCK-8、EdU细胞增殖实验、克隆形成实验检测细胞活力、增殖及克隆情况,划痕实验、Trnaswell小室侵袭实验检测细胞迁移、侵袭情况,F-actin染色观察细胞骨架形态.通过RNA-seq分析高剂量HDE干预对微小RNA(miRNA)的影响,通过实时荧光定量聚合酶链式反应法(qRT-PCR)验证HDE对HCC-7721细胞中差异miRNA表达的影响.在高剂量HDE干预的同时过表达或低表达miR-340,观察肝癌细胞活力、增殖、克隆、迁移和侵袭能力的变化.结果 HDE能呈剂量依赖地抑制肝癌细胞HCC-7721、LM3细胞活力、增殖、克隆、迁移和侵袭能力,比较差异均有统计学意义(P<0.05).F-actin染色发现,NC组细胞的丝状伪足及片状伪足较多;HDE-L组和HDE-H组细胞形态出现皱缩,细胞间连接减少,丝状伪足及片状伪足也明显减少,同时2组F-actin荧光强度较对照组均降低(P<0.05),且HDE-H组低于HDE-L组(P<0.05).RNA-seq显示,HDE干预HCC-7721细胞后,共检测到1769个差异表达基因,其中miR-340表达变化最显著,较对照细胞上调12.64倍,且qRT-PCR也证实,HDE能呈剂量依赖地上调HCC-7721细胞中miR-340的表达(P<0.05).在高剂量HDE干预的同时过表达miR-340后,肝癌细胞HCC-7721、LM3的细胞活力、增殖、克隆、迁移和侵袭能力均下降(P<0.05),低表达miR-340后,肝癌细胞HCC-7721、LM3的细胞活力、增殖、克隆、迁移和侵袭能力均增强(P<0.05).结论 HDE通过上调miR-340的表达,进而抑制肝癌细胞HCC-7721、LM3增殖、迁移及侵袭能力.
Effect of Hedyotisdiffusa extract on the proliferation,migration,and invasion of hepatocellular carcinoma cells by regulating MiR-340
Objective To explore the effect of Hedyotisdiffusa extract(HDE)on the proliferation,migration,and invasion of liver cancer cells and the underlying mechanism.Methods Liver cancer cells HCC-7721 and LM3 induced with blank control,and low-dose and high-dose HDE for 48 h.Cell viability,proliferation and colony were assessed by cell counting kit-8(CCK-8)assay,EdU assay and colony formation,respectively.Cell migration and invasion were assessed by wound healing assay and Trasnwell assay,respectively.Cell cytoskeleton was observed by immunostaining of F-actin.RNA-seq was performed to identify differentially expressed genes in liver cancer cells induced with high-dose HDE,and verified in HCC-7721 cells by quantitative reverse-transcription polymerase chain reaction(qRT-PCR).Changes in the viability,proliferation,colony,migration and invasion of liver cancer cells induced with high-dose HDE and overexpression/knockdown of miR-340 were detected.Results HDE dose-dependently inhibited the viability,proliferation,colony,migration and invasion of HCC-7721 and LM3 cells(P<0.05).F-actin staining found that liver cancer cells induced with blank control had more filopodia and lamellipodia,while those induced with low-dose and high-dose HDE showed shrinkage,reduced intercellular connections,and filopodia and lamellipodia.The fluorescence intensity of F-actin was significantly lower in liver cancer cells induced with low-dose and high-dose HDE than those of controls,which was significantly lower in the high-dose HDE group than the low-dose HDE group(P<0.05).RNA-seq showed 1 769 differentially expressed genes in HCC-7721 cells induced with high-dose HDE,among which miR-340 was the mostly upregulated for 12.64 times.qRT-PCR also confirmed that HDE dose-dependently upregulated miR-340 in HCC-7721 cells(P<0.05).High-dose HDE induction and overexpression of miR-340 significantly inhibited the viability,proliferation,colony,migration and invasion of HCC-7721 and LM3 cells(P<0.05).Knockdown of miR-340 significantly promoted the viability,proliferation,colony,migration and invasion of HCC-7721 and LM3 cells(P<0.05).Conclusion HDE upregulats miR-340,thereby inhibiting the cell viability,migration,and invasion abilities of liver cancer cells HCC-7721 and LM3.

Hepatocellular carcinomaHedyotisdiffusaMiR-340Neoplasm metastasisNeoplasm invasivenessCell proliferationPharmacologic mechanisms of action(traditional Chinese medicine)

彭亚楠、单丽芳、姜俊玲、唐晓冰、康国娇、罗克胜、周葵、赵强

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云南省普洱市中医医院药剂科,云南 普洱 665000

云南省澜沧县中医医院药剂科,云南 澜沧 665600

云南省普洱市民族传统医药研究所制剂室,云南 普洱 665000

肝癌,肝细胞 白花蛇舌草 miR-340 肿瘤转移 肿瘤浸润 细胞增殖 药理作用分子作用机制(中药)

2024

河北中医
河北省医学情报研究所,河北省中医药学会

河北中医

CSTPCD
影响因子:0.951
ISSN:1002-2619
年,卷(期):2024.46(12)