Comprehensive circular RNA expression profiles and the tumor-suppressive function of circHIPK3 in ovarian cancer
Comprehensive circular RNA expression profiles and the tumor-suppressive function of circHIPK3 in ovarian cancer
复制标题
卵巢癌中circHIPK3的综合环状RNA表达谱和肿瘤抑制功能
DOI：
10.1016/j.biocel.2019.04.011
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发表时间：
2019-07-01
影响因子：
4
通讯作者：
Jia, Xuemei
中科院分区：
文献类型：
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作者：
Teng, Fang;Xu, Juan;Jia, Xuemei
背景资料：随着新一代测序技术的发展，人们发现了数以千计的环状RNA（circular RNA，circRNA），许多circRNA已被证实在肿瘤发生中起着重要作用。然而，circRNA是否参与卵巢癌的发生和发展仍有待澄清。方法：我们分析了circRNA表达谱在上皮性卵巢癌（EOC）和正常卵巢组织（NOT）使用NGS和验证六个随机选择的circRNA通过定量实时PCR（qRT-PCR），逆转录PCR（RT-PCR）和桑格测序后RNase处理。CircHIPK 3是我们测序数据中最丰富的circRNA，它被siRNA进一步敲低。在circHIPK 3被有效沉默后，通过细胞计数试剂盒8（CCK 8）、伤口愈合、transwell和流式细胞术分析circHIPK 3在卵巢癌细胞和正常卵巢上皮细胞的增殖、侵袭、迁移和凋亡中的作用。共发现7333个circRNA，其中4505个（61.43%）是新发现的，2431个显著上调，3120个显著下调。在18例EOC和18例NOT中检测了6个随机选择的差异表达的circRNA。此外，RT-PCR和桑格测序的结果后RNase处理证实了头到尾回剪接。circHIPK 3基因沉默促进卵巢癌细胞（A2780和SKOV 3）和正常卵巢上皮细胞（IOSE 80）的增殖、迁移和侵袭，并抑制凋亡。结论：我们鉴定了卵巢癌组织中的circRNA表达谱，并进一步验证了随机选择的6个差异表达circRNA的存在和表达。此外，我们还发现circHIPK 3是卵巢癌进展的重要调节因子。
Background: With the development of next-generation sequencing (NGS), thousands of circular RNAs (circRNAs) have been found. Many circRNAs have been verified to play vital roles in carcinogenesis. However, whether circRNAs engage in the development and progression of ovarian cancer remains to be clarified.Methods: We analyzed circRNA expression profiling in epithelial ovarian cancer (EOC) and normal ovarian tissues (NOT) using NGS and validated six randomly selected circRNAs via quantitative real-time-PCR (qRT-PCR), reverse-transcription PCR (RT-PCR) and Sanger sequencing after RNase treatment. CircHIPK3, the most abundant circRNA in our sequencing data, was further knocked down by siRNA. The circHIPK3 function in proliferation, invasion, migration and apoptosis of ovarian cancer cells and normal ovarian epithelial cells was analyzed via cell counting-kit 8 (CCK8), wound healing, transwell and flow cytometry analyses after circHIPK3 was efficiently silenced.Results: Altogether, we found 7333 circRNAs, of which 4505 (61.43%) were newly identified, 2431 were significantly upregulated and 3120 were remarkably downregulated. Six randomly selected differentially expressed circRNAs were examined in 18 EOC and 18 NOT. Furthermore, the results of RT-PCR and Sanger sequencing after RNase treatment confirmed head-to-tail back-splicing. Silencing of circHIPK3 promoted proliferation, migration, and invasion and inhibited apoptosis of ovarian cancer cells (A2780 and SKOV3) and normal ovarian epithelial cells (IOSE80). Additionally, the circHIPK3-miRNA-mRNA axis was predicted as the possible mechanism using bioinformatic approaches.Conclusions: We identified the circRNA expression profile in ovarian cancer tissues and further verified the existence and expression of six randomly selected differentially expressed circRNAs. Besides, we also found that circHIPK3 is an important regulator of ovarian cancer progression.