Hsa_circ_0003998 通过海绵吸附 miR-143-3p 和 PCBP1 促进肝癌的上皮间质转化。
Hsa_circ_0003998 promotes epithelial to mesenchymal transition of hepatocellular carcinoma by sponging miR-143-3p and PCBP1.
机构信息
Department of Oncology, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, 1111, Xianxia Road, Shanghai, 200336, China.
Third Department of Hepatic Surgery, Eastern Hepatobiliary Surgery Hospital, Naval military Medical University, Shanghai, China.
出版信息
J Exp Clin Cancer Res. 2020 Jun 17;39(1):114. doi: 10.1186/s13046-020-01576-0.
BACKGROUND
Circular RNAs (circRNAs) play a critical regulatory role in cancer progression. However, the underlying mechanisms of circRNAs in hepatocellular carcinoma (HCC) metastasis remain mostly unknown.
METHODS
Has_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.
RESULTS
Compared 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.
CONCLUSION
Circ0003998 promotes EMT of HCC by circ0003998/miR-143-3p/FOSL2 axis and circ0003998 /PCBP1/CD44v6 axis.
背景
环状 RNA（circRNAs）在癌症进展中发挥着关键的调控作用。然而，circRNAs 在肝细胞癌（HCC）转移中的潜在机制在很大程度上仍不清楚。
方法
通过对伴有/不伴有门静脉癌栓（PVTT）转移的 HCC 患者的 RNA 测序，鉴定出 Has_circ_0003998（circ0003998）。通过原位杂交组织微阵列（ISH-TMA）和 25 例伴有 PVTT 转移的 HCC 患者的 qRT-PCR 进一步检测 circ0003998 的表达水平。此外，还招募了 25 例伴有 PVTT 转移的 HCC 患者和 50 例不伴有 PVTT 转移的 HCC 患者，分析 circ0003998 表达与 HCC 临床特征的相关性。利用 Transwell、迁移和 CCK8 测定以及裸鼠肺或肝转移模型，评估 circ0003998 在 HCC 上皮间质转化（EMT）中的作用。通过双荧光素酶报告基因检测、核质分离、荧光原位杂交、RNA 下拉、微 RNA 序列、western blot 和 RNA 免疫沉淀确定 circ0003998 在 miR-143-3p 和 PCBP1 中的调控机制。
结果
与相邻正常肝组织（ANL）相比，circ0003998 在 PVTT 组织和 HCC 组织中的表达明显上调，其表达与 HCC 患者侵袭性特征相关。进一步的实验表明，circ0003998 可在体外和体内促进 HCC 的 EMT。机制上，我们的数据表明，circ0003998 可能作为 microRNA-143-3p 的 ceRNA（竞争性内源性 RNA），解除其对 EMT 相关刺激因子 FOSL2 的抑制作用；同时，circ0003998 可与 PCBP1-多聚（rC）结合蛋白 1（PCBP1）结合，增加 EMT 相关基因 CD44v6 的表达水平。
结论
circ0003998 通过 circ0003998/miR-143-3p/FOSL2 轴和 circ0003998/PCBP1/CD44v6 轴促进 HCC 的 EMT。
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