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山东大学学报 (医学版) ›› 2023, Vol. 61 ›› Issue (2): 78-87.doi: 10.6040/j.issn.1671-7554.0.2022.1273

• 临床医学 • 上一篇    下一篇

环状RNA hsa_circ_0008591对乳腺癌细胞生物学行为的影响

董相君1,李娟1,孔雪1,李培龙1,赵文静2,梁怡然3,王丽丽4,杜鲁涛1,王传新1   

  1. 1.山东大学第二医院检验医学中心, 山东 济南 250033;2.山东大学齐鲁医院科研处生物样本资源库, 山东 济南 250012;3.山东大学齐鲁医院乳腺外科, 山东 济南 250012;4.山东大学齐鲁医院检验医学中心, 山东 济南 250012
  • 发布日期:2023-02-17
  • 通讯作者: 王传新. E-mail:cxwang@sdu.edu.cn
  • 基金资助:
    国家自然科学基金(82002229);山东省自然科学基金(ZR2019PH074)

Effects of circular RNA hsa_circ_0008591 on tumor biological behavior of breast cancer cells

DONG Xiangjun1, LI Juan1, KONG Xue1, LI Peilong1, ZHAO Wenjing2, LIANG Yiran3, WANG Lili4, DU Lutao1, WANG Chuanxin1   

  1. 1. Department of Clinical Laboratory, The Second Hospital of Shandong University, Jinan 250033, Shandong, China;
    2. Pathology Tissue Bank, Department of Scientific Research, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China;
    3. Department of Breast Surgery, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China;
    4. Department of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
  • Published:2023-02-17

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摘要： 目的 探讨环状RNA(circRNA)hsa_circ_0008591在乳腺癌中的表达及其对乳腺癌细胞生物学行为的影响。 方法 运用高通量芯片数据分析筛选在乳腺癌组织中差异表达的circRNAs,确定hsa_circ_0008591为研究对象,并通过Sanger测序、核糖核酸酶R(RNaseR)处理等对其进行表征。利用实时荧光定量PCR(qRT-PCR)检测hsa_circ_0008591在乳腺癌组织和细胞中的表达;进一步将质粒或小干扰RNA(siRNA)转染至乳腺癌细胞,采用实时无标记细胞分析技术(RTCA)、CCK-8增殖实验、EdU增殖实验和平板克隆形成实验检测hsa_circ_0008591对细胞增殖能力的影响。运用TargetScan等数据库预测互相作用的微小RNA(miRNA)和蛋白,同时进行功能富集分析;并运用Cytoscape软件绘制竞争性内源RNA(ceRNA)调控网络。 结果 与癌旁组织比较,hsa_circ_0008591在乳腺癌组织中呈低表达(P=0.007 6);与人正常乳腺上皮细胞比较,hsa_circ_0008591在6种乳腺癌细胞株中的表达水平明显下调(P<0.001)。体外功能实验结果表明,过表达hsa_circ_0008591可抑制MDA-MB-231(PRTCA<0.001,PEdU=0.000 6,P克隆=0.001 0)、MCF-7(PRTCA<0.001,PEdU=0.001 7,P克隆<0.001)细胞的增殖能力,而敲减hsa_circ_0008591可促进SK-BR-3(Pcck-8<0.001,PEdU<0.001,P克隆=0.003 4)细胞的增殖能力。下游靶点和通路富集分析结果显示,hsa_circ_0008591可能通过ceRNA机制或与RNA结合蛋白(RBP)相互作用抑制乳腺癌的进展。 结论 circRNA hsa_circ_0008591可抑制乳腺癌细胞增殖,有望成为乳腺癌治疗的有效干预靶点。

关键词: 环状RNA, Hsa_circ_0008591, 乳腺癌, 增殖, 生物学行为

Abstract: Objective To investigate the expression of circular RNA(circRNA)hsa_circ_0008591 and its effects on the biological behavior of breast cancer(BC)cells. Methods The differentially expressed circRNAs in BC tissues were screened using high-throughput circRNA microarray, and hsa_circ_0008591 was selected as a candidate, which was characterized by Sanger sequencing and ribonuclease R(RNaseR)treatment. Quantitative real-time PCR(qRT-PCR)was performed to detect the expression of hsa_circ_0008591 in BC tissues and cells. Furthermore, BC cells were transfected with plasmid or small interfering RNA(siRNA)and the effects of hsa_circ_0008591 on cell proliferation were detected with real-time cellular analysis(RTCA), cell counting kit-8(CCK-8)assay, 5-ethynyl-2'-deoxyuridine(EdU)assay, and colony formation assay. The interacting microRNAs(miRNAs)and proteins were predicted by databases such as TargetScan, and function enrichment analysis were performed. A regulatory network of ceRNA was constructed with Cytoscape software. Results Hsa_circ_0008591 was significantly downregulated in BC tissues(P=0.007 6). Compared with normal breast epithelial cells, six BC cell lines had a lower expression of hsa_circ_0008591(P<0.001). In vitro functional experiments showed that overexpression of hsa_circ_0008591 inhibited the proliferation of MDA-MB-231(PRTCA<0.001, PEdU=0.000 6, Pcolony=0.001 0)and MCF-7 cells(PRTCA<0.001, PEdU=0.001 7, Pcolony<0.001), while the knockdown of hsa_circ_0008591 promoted the proliferation of SK-BR-3 cells(PRTCA<0.001, PEdU<0.001, Pcolony=0.003 4). The downstream targets, gene ontology(GO)analysis, and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment indicated that hsa_circ_0008591 inhibited the progression of BC through ceRNA mechanism or interaction with RNA-binding protein(RBP). Conclusion Circular RNA hsa_circ_0008591 can inhibit the proliferation of BC cells and is expected to be an effective intervention target for the treatment of BC.

Key words: Circular RNA, Hsa_circ_0008591, Breast cancer, Proliferation, Biological behavior

中图分类号: 

  • R737.9
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