中国癌症杂志 ›› 2019, Vol. 29 ›› Issue (11): 845-854.doi: 10.19401/j.cnki.1007-3639.2019.11.002

• 论著 • 上一篇    下一篇

环状RNA hsa_circ_0005320在三阴性乳腺癌中的表达及其对细胞增殖的影响

郑夏颖,陈俊霞   

  1. 重庆医科大学细胞生物学与遗传学教研室,重庆 400016
  • 出版日期:2019-11-30 发布日期:2019-12-09
  • 通信作者: 陈俊霞 E-mail: chjunxia@126.com
  • 基金资助:
    国家自然科学基金(81672536)。

Expression of circRNA hsa_circ_0005320 in triple-negative breast cancer and its effect on cell proliferation

ZHENG Xiaying, CHEN Junxia   

  1. Department of Cell Biology and Genetics, Chongqing Medical University, Chongqing 400016, China
  • Published:2019-11-30 Online:2019-12-09
  • Contact: CHEN Junxia E-mail: chjunxia@126.com

摘要: 背景与目的:环状RNA(circular RNA,circRNA)作为新近发现的具有重要调节潜能的非编码RNA,与多种肿瘤的发生、发展密切相关,但在三阴性乳腺癌(triple-negative breast cancer,TNBC)中罕见报道。探讨hsa_circ_0005320在TNBC中的表达变化及其对细胞增殖的影响。方法:通过RNA测序(RNA sequencing,RNA-Seq)分析4对于重庆医科大学附属第一医院行手术切除的TNBC组织和癌旁组织,采用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)验证20对TNBC组织和癌旁组织以及正常人乳腺上皮细胞MCF-10A和TNBC细胞MDA-MB-231、BT-549中hsa_circ_0005320的表达,并通过荧光原位杂交(fluorescence in situ hybridization,FISH)实验进一步检测其在TNBC中的表达。采用RTFQ-PCR检测沉默hsa_circ_0005320后TNBC细胞中hsa_circ_0005320的表达;采用细胞计数试剂盒(cell counting kit-8,CCK-8)和EdU实验检测细胞增殖;采用流式细胞术、Hoechst 33342、Tunel实验检测细胞凋亡;采用流式细胞术检测细胞周期;采用蛋白质印迹法(Western blot)检测LIF-STAT3通路相关蛋白的表达情况。结果:hsa_circ_0005320在TNBC组织和细胞中显著高表达;在TNBC细胞中沉默hsa_circ_0005320后其表达量显著降低,细胞增殖能力下降,促进细胞凋亡,导致细胞周期阻滞在G1期,且LIF、P-STAT3蛋白表达水平明显下降。结论:hsa_circ_0005320在TNBC中高表达,沉默hsa_circ_0005320可显著抑制TNBC细胞的增殖,诱导细胞凋亡。

关键词: 环状RNA, 三阴性乳腺癌, hsa_circ_0005320, 增殖

Abstract: Background and purpose: Circular RNA (circRNA), as a newly discovered non-coding RNA with important regulatory potential, is closely related to the occurrence and development of various tumors, but it is rarely reported in triple-negative breast cancer (TNBC). The aim of the present study was to investigate the expression of circRNA hsa_circ_0005320 in TNBC and its effect on the proliferation of TNBC. Methods: RNA sequencing (RNA-seq) was adopted to analyze 4 pairs of TNBC tissues and adjacent non-cancerous tissues who received surgical resection in the First Affiliated Hospital of Chongqing Medical University. Using real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR), we detected the relative expression levels of circRNA hsa_circ_0005320 in 20 pairs of TNBC tissues and adjacent non-cancerous tissues, normal breast epithelial cell MCF-10A and TNBC cell MDA-MB-231 and BT-549, and further tested the expression by fluorescence in situ hybridization (FISH) assay. After silencing hsa_circ_0005320, the relative expression level of hsa_circ_0005320 was detected by RTFQ-PCR. The proliferation of TNBC cells was determined by cell counting kit-8 (CCK-8) and EdU assays. Flow cytometry, Hoechst 33342 and TUNEL assay were employed to detect cell apoptosis, respectively. Cell cycle distribution was analyzed by flow cytometry. Western blot was used to detect the expression levels of proteins related to the LIF-STAT3 pathway. Results: circRNA hsa_circ_0005320 was highly expressed in TNBC tissues and cells. After silencing hsa_circ_0005320 in TNBC cells, its expression was significantly reduced, the ability of cell proliferation was decreased, cell apoptosis was promoted, leading to cell cycle arrest in G1 phase, and lower protein levels BTof LIF and p-STAT3 were observed clearly. Conclusion: CircRNA hsa_circ_0005320 is highly expressed in TNBC, and silencing hsa_circ_0005320 can significantly inhibit the proliferation of TNBC cells and induce apoptosis.

Key words: Circular RNA, Triple-negative breast cancer, hsa_circ_0005320, Proliferation