Hsa_circ_0003998 promotes epithelial to mesenchymal transition of hepatocellular carcinoma by sponging miR-143-3p and PCBP1
Hsa_circ_0003998 promotes epithelial to mesenchymal transition of hepatocellular carcinoma by sponging miR-143-3p and PCBP1
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Hsa_circ_0003998 通过海绵 miR-143-3p 和 PCBP1 促进肝细胞癌上皮向间质转化
DOI：
10.1186/s13046-020-01576-0
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发表时间：
2020-06-17
影响因子：
11.3
通讯作者：
Yan, Hong-li
中科院分区：
文献类型：
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作者：
Song, Li-na;Qiao, Guang-lei;Yan, Hong-li
研究背景环状RNA（CircRNA）在肿瘤的发生、发展过程中起着重要的调控作用。方法采用RNA测序技术，在肝癌伴或不伴门静脉癌栓（PVTT）转移患者中检测到一个名为Has_circ_0003998（circ 0003998）的基因。采用组织芯片原位杂交（ISH-TMA）和qRT-PCR检测25例肝癌伴PVTT转移患者中circ 0003998的表达水平。同时选取25例有PVTT转移的HCC患者和50例无PVTT转移的HCC患者，分析circ 0003998表达与HCC临床特征的相关性。采用Transwell、迁移和CCK 8试验以及裸鼠肺或肝转移模型来评价circ 0003998在HCC上皮间质转化（EMT）中的作用。结果与癌旁正常肝组织（ANL）相比，circ 0003998在PVTT组织和HCC组织中的表达显著上调，与癌旁正常肝组织（ANL）相比，circ 0003998在PVTT组织中的表达显著上调，与癌旁正常肝组织相比，circ 0003998在PVTT组织中的表达显著上调，与癌旁正常肝组织相比，circ 0003998在PVTT组织中的表达显著上调，与癌旁正常肝组织相比，circ 0003998在PVTT组织中的表达显著上调。其表达与HCC患者的侵袭性特征相关。进一步的试验表明，circ 0003998在体外和体内均促进HCC的EMT。从机制上讲，我们的数据表明，circ 0003998可能作为一种ceRNA，microRNA-143- 3 p（竞争性内源性RNA）对EMT相关刺激因子FOSL 2的抑制作用;同时，circ 0003998可与PCBP 1-poly（rC）binding protein 1（PCBP 1）结合，提高EMT相关基因的表达水平，结论Circ 0003998通过circ 0003998/miR-143- 3 p/FOSL 2轴和circ 0003998/PCBP 1/CD 44 v6轴促进HCC的EMT。
BackgroundCircular RNAs (circRNAs) play a critical regulatory role in cancer progression. However, the underlying mechanisms of circRNAs in hepatocellular carcinoma (HCC) metastasis remain mostly unknown.MethodsHas_circ_0003998 (circ0003998) was identified by RNAs sequencing in HCC patients with /without portal vein tumor thrombus (PVTT) metastasis. The expression level of circ0003998 was further detected by in situ hybridization on tissues microarray (ISH-TMA) and qRT-PCR in 25 HCC patients with PVTT metastasis. Moreover, the 25 HCC patients with PVTT metastasis and 50 HCC patients without PVTT metastasis were recruited together to analyze the correlation between circ0003998 expression and HCC clinical characteristics. Transwell, migration and CCK8 assays, as well as nude mice model of lung or liver metastasis were used to evaluate the role of circ0003998 in epithelial to mesenchymal transition (EMT) in HCC. The regulatory mechanisms of circ0003998 in miR-143-3p and PCBP1 were determined by dual-luciferase reporter assay, nuclear-cytoplasmic fractionation, fluorescent in situ hybridization, RNA pull- down, microRNA sequence, western blot and RNA immunoprecipitation.ResultsCompared with adjacent normal liver tissues (ANL), circ0003998 expression was significantly upregulated in PVTT tissues and HCC tissues, and its expression correlates with the aggressive characteristics of HCC patients. Further assays suggested that circ0003998 promoted EMT of HCC both in vitro and in vivo. Mechanistically, our data indicated that circ0003998 may act as a ceRNA (competing endogenous RNA) of microRNA-143-3p to relieve the repressive effect on EMT-related stimulator, FOSL2; meanwhile, circ0003998 could bind with PCBP1-poly(rC) binding protein 1 (PCBP1) to increase the expression level of EMT-related genes, CD44v6.ConclusionCirc0003998 promotes EMT of HCC by circ0003998/miR-143-3p/FOSL2 axis and circ0003998 /PCBP1/CD44v6 axis.