Tu Qiang, You Xiaoxiang, He Jun, Hu Xuguang, Xie Changji, Xu Guohui
Department of Hepatobiliary Oncology Surgery, Jiangxi Cancer Hospital of Nanchang University, Nanchang, People's Republic of China.
Department of Oncology Interventional, Jiangxi Cancer Hospital of Nanchang University, Nanchang, People's Republic of China.
Cancer Manag Res. 2021 Oct 13;13:7859-7870. doi: 10.2147/CMAR.S315894. eCollection 2021.
Hepatocellular carcinoma (HCC) is one most common cancer types among gastrointestinal cancer over the world, while its underlying mechanisms remain unclear. CircRNA has been revealed to participate in multiple biological functions and contribute to various diseases' progression.
Bioinformatic analysis of the differently expressed circRNAs in the HCC tissues, then verified by real-time quantitative PCR (RT-qPCR) assay. We found that circ-0003006 was upregulated in the HCC tissues, the cell fractionation assay and RNA fluorescence in situ hybridization (FISH) were performed to confirm the cell location of circ-0003006. shRNA silence assay was used to knock down the expression of circ-0003006 in the HCC cells.
Cell account kit 8 (CCK-8) and transwell assay were revealed that circ-0003006 knockdown inhibited the proliferation and metastasis in HCC cells. The target miR‑542‑3p and target gene HIF-1A were predicted by bioinformatics analysis, then verified through biotinylated RNA pull-down and dual-luciferase reporter assays. The mechanism, circ-0003006, probably acted as a sponge of miR‑542‑3p and regulated HIF-1A levels in hepatocellular carcinoma cells. Moreover, HIF-1A overexpression abolished the effect of circ-0003006 inhibition on the progression of hepatocellular carcinoma cells. The subcutaneous tumor formation experiment indicated that circ-0003006 knockdown inhibited the HCC cell growth in vivo.
Circ-0003006 was demonstrated to promote HCC progression in vitro and in vivo by sponging miR‑542‑3p to release the inhibition on HIF-1A.
肝细胞癌（HCC）是全球胃肠道癌症中最常见的癌症类型之一，但其潜在机制仍不清楚。环状RNA（circRNA）已被揭示参与多种生物学功能，并促进各种疾病的进展。
对HCC组织中差异表达的circRNA进行生物信息学分析，然后通过实时定量PCR（RT-qPCR）检测进行验证。我们发现circ-0003006在HCC组织中上调，进行细胞分级分离实验和RNA荧光原位杂交（FISH）以确认circ-0003006的细胞定位。使用短发夹RNA（shRNA）沉默实验来敲低HCC细胞中circ-0003006的表达。
细胞计数试剂盒8（CCK-8）和Transwell实验表明，circ-0003006敲低抑制了HCC细胞的增殖和转移。通过生物信息学分析预测了靶标miR-542-3p和靶基因缺氧诱导因子-1α（HIF-1A），然后通过生物素化RNA下拉实验和双荧光素酶报告基因实验进行验证。机制上，circ-0003006可能作为miR-542-3p的海绵，调节肝癌细胞中HIF-1A的水平。此外，HIF-1A过表达消除了circ-0003006抑制对肝癌细胞进展的影响。皮下肿瘤形成实验表明，circ-0003006敲低在体内抑制了HCC细胞生长。
Circ-0003006被证明通过海绵吸附miR-542-3p以解除对HIF-1A的抑制，从而在体外和体内促进HCC进展。
