医学分子生物学杂志 ›› 2024, Vol. 21 ›› Issue (5): 405-412.doi: 10.3870/j.issn.1672-8009.2024.05.003

• 论著 • 上一篇    下一篇

miR-542-5p 通过靶向结合 ENSA 抑制肝细胞癌增殖迁移和侵袭 #br#

  

  1. 1武汉市第九医院检验科 武汉市, 430081 武汉钢铁 (集团) 公司第二职工医院2肿瘤科,3检验科 武汉市, 430085
  • 出版日期:2024-09-30 发布日期:2024-10-25
  • 基金资助:
    湖北省自然科学基金 (No. 2017CFL083)

miR-542-5p Inhibits Proliferation, Migration and Invasion of Hepatocellular Carcinoma by Targeting ENSA #br#

  1. 1 Department of Clinical Laboratory, Wuhan Ninth Hospital, Wuhan, 430081, China 2 Department of Oncology,3Department of Clinical Laboratory, the Second Staff Hospital of Wuhan Iron and Steel (Group) Company, Wuhan, 430085, China
  • Online:2024-09-30 Published:2024-10-25

摘要: 目的 探讨 miR-542-5p 靶向结合 ENSA 对肝细胞癌增殖迁移和侵袭的影响方法 通过GSE36915 数据矩阵分析 miR-542-5p 在肝癌组织和癌旁组织表达情况, 构建 miR-542-5p 过表达肝癌细胞HepG2 Hep3B 细胞系并使用 RT-PCR 验证, 采用 CCK-8、 划痕、 Transwell 法检测细胞增殖迁移和侵袭能力; 使用 miR-542-5p 过表达 Hep3B 细胞系建立体内裸鼠移植瘤, 观察移植瘤体积变化和重量变化, 使用RT-PCR 检测移植瘤 miR-542-5p 水平通过 GEO2R 分析 GSE101728、 GSE7642 的差异基因, 使用 miRWalk在线预测 miR-542-5p 的靶基因, 筛选符合以上条件的基因得到 ENSA; 通过 TCGA 肝癌数据分析 ENSA 在肝癌组织和癌旁组织的表达情况, RT-PCR 检测 miR-542-5p 过表达 HepG2 Hep3B 细胞 ENSA 水平, 并在miR-542-5p 过表达 HepG2 Hep3B 细胞系中转染 ENSA 过表达质粒, 使用上述相同方法检测细胞增殖迁移和侵袭能力结果 miR-542-5p 在肝癌组织中表达水平显著低于癌旁组织 ( P< 0. 01), miR-542-5p 过表达 HepG2 Hep3B 细胞系的光密度 (A ) 和相对细胞侵袭数均明显低于其阴性对照 ( miR-NC) ( P< 0. 01), 而划痕宽度明显高于其 miR-NC (P< 0. 01); 裸鼠移植瘤实验结果显示 miR-542-5p 过表达 Hep3B 细胞瘤的体积和重量明显低于 miR-NC ( P< 0. 01)。 ENSA 在肝癌组织中表达水平显著高于癌旁组织 ( P < 0. 01) ENSA 促进肝癌进展; miR-542-5p 过表达 HepG2 Hep3B 细胞系中 ENSA 蛋白表达显著降低于其阴性对照 (miR-NC) (P< 0. 01); miR-542-5p + ENSA 双过表达 HepG2 Hep3B 细胞系的 A 值和细胞侵袭数均明显高于其 miR-542-5p 过表达细胞 (P< 0. 01), 而划痕宽度明显低于其 miR-542-5p 过表达细胞 ( P< 0. 01)。 结论 miR-542-5p 可通过靶向抑制 ENSA 表达抑制肝细胞癌细胞增殖迁移和侵袭

关键词: 肝细胞癌, miR-542-5p, ENSA, 细胞增殖, 细胞迁移, 细胞侵袭

Abstract: Objective To explore the effect of miR-542-5p on proliferation, migration and invasion of hepatocellular carcinoma (HCC) by targeting ENSA. Methods The expression of miR-542-5p in HCC tissues and para-carcinoma tissues was analyzed by GSE36915 data matrix. miR-542- 5p overexpression were performed in the HCC cell lines HepG2 and Hep3B and verified by RTPCR. The proliferation, migration and invasion of cells were detected by CCK-8, wound healing and Transwell assays. miR-542-5p overexpressed Hep3B cells were used to establish the transplanted tumor model in nude mice. The volume and weight of transplanted tumors were measured. The miR- 542-5p expression level in transplanted tumors was detected by RT-PCR. The differential expression genes of GSE101728 and GSE7642 were analyzed by GEO2R, and the target genes of miR-542-5pwere predicted by miRWalk, ENSA gene was then screened out. The expression level of ENSA inHCC tissues and para-carcinoma tissues was analyzed by TCGA HCC data. The expression level of ENSA in miR-542-5p overexpressed HepG2 and Hep3B cells was detected by RT-PCR. The cell proliferation, migration and invasion were detected in miR-542-5p overexpressed HepG2 and Hep3Bcells after transfected with ENSA overexpression plasmid. Results The expression level of miR-542-5p in the HCC tissues was significantly lower than that in the para-carcinoma tissues (P < 0. 01).The proliferation rate, migration rate and relative number of invasion cells in the miR-542-5p overexpressed cells were significantly lower than those in miR-NC overexpressed cells (P < 0. 01). The results ofin vivoexperiments showed that the volume and weight of tumors in the miR-542-5p overexpression group were significantly lower than those in the miR-NC overexpression group (P< 0. 01).The expression level of ENSA in the HCC tissues was significantly higher than that in the para-carcinoma tissues (P< 0. 01), and ENSA could promote HCC progression. The protein expression levelof ENSA in the miR-542-5p overexpressed cells were significantly lower than that in the miR-NCoverexpressed cells (P< 0. 01). The proliferation rate, migration rate and relative number of invasion cells in cells with miR-542-5p + ENSA overexpression were significantly higher than those incells with miR-542-5p overexpression (P< 0. 01). Conclusion miR-542-5p can inhibit the proliferation, migration and invasion of HCC cells by inhibiting ENSA.

Key words:

hepatocellular carcinoma, miR-542-5p, ENSA, cell proliferation, cell migration, cell invasion

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