Yu Yanhui, Bian Lijuan, Liu Renfei, Wang Yitong, Xiao Xia
Department of Clinical Laboratory, The Second Hospital of Jilin University, No. 218ZiQiang Street, Changchun, 130001, Jilin, China.
Department of Cataract, Eye Center, The Second Hospital of Jilin University, Changchun, Jilin, China.
Cancer Cell Int. 2021 Jan 6;21(1):10. doi: 10.1186/s12935-020-01695-w.
Circular RNA hsa_circ_0061395 (circ_0061395) has been reported to accelerate the advancement of hepatocellular carcinoma (HCC). However, the regulatory mechanism by which circ_0061395 modulates the progression of HCC is unclear.
The morphology and size of exosomes were analyzed by transmission electron microscope (TEM) and nanoparticle-tracking analysis (NTA). Protein levels were detected by western blotting. Expression levels of circ_0061395, microRNA (miR)-877-5p, and phosphoinositide-3-kinase regulatory subunit 3 (PIK3R3) mRNA were assessed by quantitative real time polymerase chain reaction (qRT-PCR). The proliferation, invasion, migration, cell cycle progression, and apoptosis were analyzed by cell counting kit-8 (CCK-8), plate clone, transwell, or flow cytometry assays. The targeting relationship between circ_0061395 or PIK3R3 and miR-877-5p was verified using the dual-luciferase reporter and/or RNA immunoprecipitation (RIP) assays. Xenograft assay was performed to confirm the biological function of circ_0061395 in HCC.
Circ_0061395 was upregulated in HCC tissues, serum, cells, and serum-derived exosomes. Circ_0061395 silencing decreased tumor growth in vivo, and induced cell cycle arrest, apoptosis, repressed proliferation, invasion, and migration of HCC cells in vitro. MiR-877-5p was downregulated while PIK3R3 was upregulated in HCC. Circ_0061395 regulated PIK3R3 expression via competitively binding to miR-877-5p. MiR-877-5p inhibitor overturned circ_0061395 knockdown-mediated influence on malignant behaviors of HCC cells. PIK3R3 overexpression reversed the suppressive influence of miR-877-5p mimic on malignant behaviors of HCC cells.
Circ_0061395 facilitated HCC progression via regulating the miR-877-5p/PIK3R3 axis, providing a new perspective on the advancement of HCC.
环状RNA hsa_circ_0061395（circ_0061395）已被报道可加速肝细胞癌（HCC）的进展。然而，circ_0061395调节HCC进展的调控机制尚不清楚。
通过透射电子显微镜（TEM）和纳米颗粒跟踪分析（NTA）分析外泌体的形态和大小。通过蛋白质印迹法检测蛋白质水平。通过定量实时聚合酶链反应（qRT-PCR）评估circ_0061395、微小RNA（miR）-877-5p和磷酸肌醇-3-激酶调节亚基3（PIK3R3）mRNA的表达水平。通过细胞计数试剂盒-8（CCK-8）、平板克隆、Transwell或流式细胞术分析评估细胞的增殖、侵袭、迁移、细胞周期进程和凋亡。使用双荧光素酶报告基因和/或RNA免疫沉淀（RIP）分析验证circ_0061395或PIK3R3与miR-877-5p之间的靶向关系。进行异种移植实验以确认circ_0061395在HCC中的生物学功能。
circ_0061395在HCC组织、血清、细胞和血清来源的外泌体中上调。circ_0061395沉默可降低体内肿瘤生长，并在体外诱导细胞周期停滞、凋亡，抑制HCC细胞的增殖、侵袭和迁移。在HCC中，miR-877-5p下调而PIK3R3上调。circ_0061395通过竞争性结合miR-877-5p来调节PIK3R3的表达。miR-877-5p抑制剂可逆转circ_0061395敲低介导的对HCC细胞恶性行为的影响。PIK3R3过表达可逆转miR-877-5p模拟物对HCC细胞恶性行为的抑制作用。
circ_0061395通过调节miR-877-5p/PIK3R3轴促进HCC进展，为HCC的进展提供了新的视角。
