Hsa_circ_001193 通过靶向 mir-496 调节鼻咽癌细胞的增殖和凋亡。
Hsa_circ_001193 regulates proliferation and apoptosis of nasopharyngeal carcinoma cells through targeting mir-496.
机构信息
Shenzhen Key Laboratory of Viral Oncology, The Clinical Innovation & Research Centre, Shenzhen Hospital, Southern Medical University, Shenzhen, Guangdong, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Mar;24(6):3069-3076. doi: 10.26355/eurrev_202003_20671.
OBJECTIVE
To explore the effects of hsa_circ_001193 on the proliferation and apoptosis of nasopharyngeal carcinoma (NPC) cells.
MATERIALS AND METHODS
The messenger ribonucleic acid (mRNA) expression level of hsa_circ_001193 in three NPC cell lines (CNE-1, SUNE-1, and HONE-1) and human normal nasopharyngeal epithelial cell line (NP69) was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). The expression of hsa_circ_001193 was silenced through transient transfection with small-interfering RNA (siRNA). Regulatory effects of hsa_circ_001193 knockdown on the proliferation and apoptosis of HONE-1 cells were determined using cell counting kit-8 (CCK-8) assay, colony formation assay, and flow cytometry. Potential miRNAs binding hsa_circ_001193 were predicted in the StarBase, which was further verified via Dual-Luciferase reporter assay and qRT-PCR. Moreover, the involvement of the predicted target miRNA in the proliferation of HONE-1 cells regulated by hsa_circ_001193 was determined by CCK-8 assay.
RESULTS
Compared with that in human normal nasopharyngeal epithelial cell line (NP69), the expression of hsa_circ_001193 was significantly upregulated in NPC cell lines (p<0.05). The results of CCK-8 assay and colony formation assay showed that knockdown of hsa_circ_001193 could significantly suppress the cell proliferation ability and colony formation ability compared with control group (p<0.05). The results of flow cytometry revealed that the apoptosis rate in hsa_circ_001193 knockdown group was remarkably higher than that in control group (p<0.05). Besides, according to the analysis of StarBase database, there were binding sites between hsa_circ_001193 and miR-496. The Dual-Luciferase reporter assay manifested that miR-496 bound hsa_circ_001193 (p<0.05).
CONCLUSIONS
Hsa_circ_001193 can serve as the miR-496 sponge, which regulates proliferation and apoptosis of NPC cells through up-regulating miR-496. Our findings provide a new therapeutic target for NPC.
目的
探讨 hsa_circ_001193 对鼻咽癌细胞增殖和凋亡的影响。
材料与方法
采用实时定量聚合酶链反应（qRT-PCR）检测三种鼻咽癌细胞系（CNE-1、SUNE-1 和 HONE-1）和人正常鼻咽上皮细胞系（NP69）中 hsa_circ_001193 的信使核糖核酸（mRNA）表达水平。通过小干扰 RNA（siRNA）瞬时转染沉默 hsa_circ_001193 的表达。通过细胞计数试剂盒-8（CCK-8）测定、集落形成测定和流式细胞术测定 hsa_circ_001193 敲低对 HONE-1 细胞增殖和凋亡的调节作用。在 StarBase 中预测与 hsa_circ_001193 结合的潜在 miRNA，进一步通过双荧光素酶报告基因测定和 qRT-PCR 进行验证。此外，通过 CCK-8 测定确定 hsa_circ_001193 调节的 HONE-1 细胞增殖所涉及的预测靶 miRNA。
结果
与正常鼻咽上皮细胞系（NP69）相比，hsa_circ_001193 在鼻咽癌细胞系中的表达显著上调（p<0.05）。CCK-8 测定和集落形成测定结果表明，与对照组相比，hsa_circ_001193 敲低组细胞增殖能力和集落形成能力明显下降（p<0.05）。流式细胞术结果显示，hsa_circ_001193 敲低组的凋亡率明显高于对照组（p<0.05）。此外，根据 StarBase 数据库的分析，hsa_circ_001193 与 miR-496 之间存在结合位点。双荧光素酶报告基因测定显示，miR-496 与 hsa_circ_001193 结合（p<0.05）。
结论
hsa_circ_001193 可以作为 miR-496 的海绵，通过上调 miR-496 调节鼻咽癌细胞的增殖和凋亡。我们的研究结果为 NPC 提供了一个新的治疗靶点。
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