Hsa_circ_0003945 promotes progression of hepatocellular carcinoma by mediating miR‐34c‐5p/LGR4/β‐catenin axis activity

Abstract

Accumulating evidence suggests that circular RNAs (circRNAs) play essential roles in regulating cancer progression, but many circRNAs in hepatocellular carcinoma (HCC) remain unknown. Dysregulated circRNAs in HCC were identified through bioinformatics analysis of Gene Expression Omnibus data sets. Quantitative real-time PCR (qRT-PCR), Sanger sequencing, RNase R digestion and actinomycin D treatment were conducted to confirm the characterization of circRNAs. CCK-8, wound-healing and Transwell assays were performed to assess the functional roles of Hsa_circ_0003945 (Circ_0003945) in HCC cell lines. Subcellular fractionation and fluorescence in situ hybridization (FISH) were performed to locate Circ_0003945 in HCC cells. Dualluciferase reporter assay was executed to verify the binding of Circ_0003945 to microRNAs (miRNAs) or the miRNAs to their target genes. In this study, we found that Circ_0003945 was upregulated in HCC tissue, and higher Circ_0003945 expression was positively correlated with tumour size and tumour stage. Furthermore, high plasma levels of circulating Circ_0003945 were confirmed in HCC patients compared with those in non-HCC groups. The functional experiments revealed that overexpression or knockdown of Circ_0003945 promoted or attenuated tumour growth and migration, respectively. Mechanistically, Circ_0003945 might exert as a miR-34c-5p sponge to upregulate the expression of leucine-rich repeat-containing G | 2219 LYU et aL.

Metformin Suppresses Hepatocellular Carcinoma through Regulating Alternative Splicing of LGR4

“…LGR4 overexpression could promote the processes of proliferation, migration, and invasion. Although the study indicated that Hsa_circ_0003945 could regulate the miR-34c-5p/LGR4/β-catenin axis to promote the progression of hepatocellular carcinoma [16], the exact role of LGR4 is still unclear. Mechanistically, LGR4 can activate wnt/β-catenin signaling in many diseases [16][17][18].…”

“…Although the study indicated that Hsa_circ_0003945 could regulate the miR-34c-5p/LGR4/β-catenin axis to promote the progression of hepatocellular carcinoma [16], the exact role of LGR4 is still unclear. Mechanistically, LGR4 can activate wnt/β-catenin signaling in many diseases [16][17][18]. So, we detected the expression of catenin through immunofluorescence.…”

Circ_0003945: an emerging biomarker and therapeutic target for human diseases

“…Ki-67 acts as specific biomarker of cellular proliferation, regulated through cell cycle-dependent transcription and protein degradation processes ( 76 , 77 ). Notably, overexpression of circ_0003945 was significantly associated with enhanced cellular of Ki-67 levels in NSCLC and HCC cells, implicating its roles in tumor cell proliferation ( 41 , 43 ). In vivo , xenograft assays in mice had verified that stable knockdown of circ_0003945 led to reduced tumor volumes and weights compared matched controls.…”

Circular RNAs in hepatocellular carcinoma: biogenesis, function, and pathology

“…Circ-102,166, a circRNA encoded by the TEX2 gene locus, inhibited HCC development by binding with miR-182 and miR-184 to regulate the expression of their downstream targets, including FOXO3a, MTSS1, SOX7, p-RB and c-MYC (Li R. et al, 2020). CircLIFR has also been identified to function as a tumor suppressor, suppressing the proliferation and invasion of HCC cells through the miR-624-5p/GSK-3β axis (Yang L. et al, 2022).…”