实用医学杂志 ›› 2022, Vol. 38 ›› Issue (9): 1088-1093.doi: 10.3969/j.issn.1006⁃5725.2022.09.009

• 基础研究 • 上一篇    下一篇

circ⁃0008583结合EIF4A3蛋白促进肝癌细胞增殖并抑制其凋亡

吴田方1 戴东2 王常元3   

  1. 1 黔南州人民医院肝胆外科(贵州黔南 558000);2 天津医科大学肿瘤医院分子影像及核医学诊疗科(天津 300000);3 贵州省人民医院肝胆外科(贵阳 550000)

  • 出版日期:2022-05-10 发布日期:2022-05-10
  • 基金资助:
    黔南民族医学高等专科学校科研骨干支撑项目(编号:QNYZ2019⁃11);天津市自然科学基金(编号:17JCYBJC2S100)

circ⁃0008583 binds to EIF4A3 protein to promote the proliferation and inhibit the apoptosis of HCC cells

WU Tianfang*,DAI Dong,WANG Changyuan.   

  1. Department of Hepatobiliary Surgery,the People′s Hospital of Qian⁃ nan,Qiannan 558000,China

  • Online:2022-05-10 Published:2022-05-10

摘要:

目的 探讨circ⁃0008583对肝癌细胞系增殖、凋亡的影响及其作用机制。方法 生物信息学 分析联合临床样本和体外细胞实验验证 circ⁃0008583 在肝癌中的表达水平。RNase R 和放线菌素 D 处理 验证 circ⁃0008583 circRNA 特性。克隆形成实验检测细胞克隆形成能力,TUNEL 染色检测细胞凋亡。 RIP 实验检测 circ⁃0008583 EIF4A3 是否结合。qRT⁃PCR Western blot 检测基因表达水平。结果 与正 常肝组织及细胞系相比,circ⁃0008583 在肝癌组织和细胞系中高表达(均 P < 0.001)。circ⁃0008583 具有环 状结构,与 DLG1 mRNA 相比,其稳定性增加(P < 0.001)。与阴性对照组相比,干扰 circ⁃0008583 表达后肝 癌细胞的克隆形成能力降低(P < 0.01),凋亡水平上升(P < 0.001)。Circular RNA Interactome 数据库和 RIP 实验证实 circ⁃0008583 能够与 EIF4A3 蛋白结合。与正常肝组织及细胞系相比,EIF4A3 在肝癌组织及细胞 中高表达(P < 0.001)。过表达 EIF4A3 能够在一定程度上逆转干扰 circ⁃0008583 表达对肝癌细胞增殖的抑 制作用以及凋亡的促进作用。结论 circ⁃0008583 在肝癌组织及细胞系中高表达,其与 EIF4A3 蛋白结合 促进肝癌细胞增殖,并抑制肝癌细胞凋亡。

关键词:

circ?0008583, EIF4A3, 肝癌, 增殖, 凋亡

Abstract:

Objective The aim of this study was to investigate the effects and mechanism of circ⁃0008583 on proliferation and apoptosis of hepatoma carcinoma cell(HCC). Methods Bioinformatics analysis combined clinical samples and in vitro cell experiments were verified the expression level of circ⁃0008583 in HCC. RNase R and actinomycin D were used to verify the circRNA properties of circ⁃0008583. Cloning formation experiment was used to test cell clone formation capacity,and TUNEL staining was used to detected cell apoptosis. We used the RIP experiment to detect whether circ ⁃ 0008583 could combined with EIF4A3. qRT ⁃ PCR and Western blot were used to detect gene expression levels. Results Compared to normal liver tissues and cell lines,circ⁃0008583 was highly expressed in HCC tissues and cell lines(P < 0.001). Circ⁃0008583 has a circular structure and its stability is increased compared with DLG1 mRNA(P < 0.001). Compared to the negative control group,the clonal formation capacity of HCC cells after interfering circ ⁃0008583 was reduced(P < 0.01),and apoptosis level was increased (P < 0.001). We use Circular RNA interactome database and RIP experiments confirm that circ ⁃ 0008583 could combine with EIF4A3 protein. Compared to normal liver tissues and cell lines,EIF4A3 was highly expressed in HCC tissues and cell line(P < 0.001). We confirmed that overexpression EIF4A3 could reverse the inhibitory effects of circ ⁃ 0008583 expression on liver cancer cell proliferation and apoptosis by rescue experiments. Conclusion Circ⁃0008583 is highly expressed in HCC tissues and cell lines,it combines with EIF4A3 protein to promote HCC cell proliferation and inhibit cell apoptosis.

Key words:

circ?0008583, EIF4A3, hepatoma carcinoma(HCC), proliferation, apoptosis ,