Bu Nan, Dong Zheng, Zhang Lingfeng, Zhu Weibo, Wei Furong, Zheng Sheng
Department of Gastroenterology, Chinese Medicine Hospital of Jiamusi City, Heilongjiang Province, Jiamusi, Heilongjiang, People's Republic of China.
Department of General Surgery, Ningbo Mingzhou Hospital, Ningbo, Zhejinag, People's Republic of China.
Cancer Manag Res. 2020 Dec 16;12:12945-12956. doi: 10.2147/CMAR.S280478. eCollection 2020.
Recent studies reported that circular RNAs (circRNAs) exert essential functions in hepatocellular carcinoma (HCC) progression. However, the expression profile and function of circular RNA PVT1 (circPVT1) in HCC are not fully addressed. Thus, we aimed to probe into the function of circPVT1 in HCC development.
The levels of circPVT1, microRNA-377 (miR-377) and transcripts encoding tripartite motif containing 23 () were determined by qRT-PCR. The stability and localization of circPVT1 were examined by RNase R digestion assay and subcellular fraction assay, respectively. Cell proliferation and apoptosis were evaluated by MTT assay and flow cytometry analysis, respectively. The relationship between miR-377 and circPVT1 or was determined by dual-luciferase reporter assay and RNA immunoprecipitation (RIP). The protein expression of was measured by Western blot. The glycolysis level was assessed by specific kits and Seahorse Extracellular Flux Analyzer XF96. The function of circPVT1 in vivo was investigated in a murine xenograft model.
CircPVT1 and levels were elevated, while miR-377 was decreased in HCC. CircPVT1 knockdown restrained proliferation and glycolysis, but enhanced apoptosis in HCC cells. CircPVT1 could bind to miR-377 and inhibition of miR-377 restored circPVT1 knockdown-mediated effect on HCC cells. was certified as a target of miR-377, and upregulation overturned the influence of miR-377 upregulation or circPVT1 silence on HCC progression. Moreover, circPVT1 knockdown restrained tumor growth in HCC in vivo.
CircPVT1 aggravated the progression of HCC by upregulating via sponging miR-377.
最近的研究报道，环状RNA（circRNAs）在肝细胞癌（HCC）进展中发挥重要作用。然而，环状RNA PVT1（circPVT1）在HCC中的表达谱和功能尚未完全明确。因此，我们旨在探究circPVT1在HCC发生发展中的作用。
通过qRT-PCR检测circPVT1、微小RNA-377（miR-377）和编码含三联基序蛋白23（ ）的转录本水平。分别通过RNase R消化试验和亚细胞分级试验检测circPVT1的稳定性和定位。分别通过MTT试验和流式细胞术分析评估细胞增殖和凋亡。通过双荧光素酶报告基因试验和RNA免疫沉淀（RIP）确定miR-377与circPVT1或 的关系。通过蛋白质免疫印迹法检测 的蛋白表达。通过特定试剂盒和Seahorse细胞外流量分析仪XF96评估糖酵解水平。在小鼠异种移植模型中研究circPVT1在体内的功能。
HCC中circPVT1和 的水平升高，而miR-377水平降低。circPVT1敲低抑制了HCC细胞的增殖和糖酵解，但增强了细胞凋亡。circPVT1可以与miR-377结合，抑制miR-377可恢复circPVT1敲低介导的对HCC细胞的作用。 被证实为miR-377的靶标，上调 可逆转miR-377上调或circPVT1沉默对HCC进展的影响。此外，circPVT1敲低在体内抑制了HCC的肿瘤生长。
circPVT1通过海绵吸附miR-377上调 ，从而促进HCC的进展。
