Bioinformatics and Experimental Insights Into miR-182, hsa_circ_0070269, and circ-102,166 as Therapeutic Targets for HCV-Associated HCC

Abstract

Aims: Hepatocellular carcinoma (HCC) is a type of malignant tumor and the sixth leading cause of death worldwide. It is caused by HBV, HCV infection, and alcohol consumption. MicroRNAs are typically small, non-coding RNAs that are involved in the regulation of mRNA expression. Recent studies revealed miRNAs' regulatory roles in liver cancer, linked to risk factors like HCV, HBV infection, alcoholism, drug use, and auto-immune hepatic disorders. Circular RNAs also belong to the class of non-coding RNAs; they act as ceRNAs to regulate miRNA expression and regulate different oncogenic pathways in HCC progression. This study aimed to check the hsa_circ_0070269, circ-102,166 (hsa_circ_0004913), and miR-182 expression in HCV induced HCC patients.

Methods: Data analysis was used to find out studies related to the role of hsa_circ_0070269, circ-102,166, and miR-182 in HCC; miR-182 targeted genes, their role in different diseases; and miR-182 interactions with hsa_circ_0070269 and circ-102,166 in the HCC. It was revealed that the hsa_circ_0070269, circ-102,166, and miR-182 correlations in HCV induced HCC have not been explored yet. Therefore, to validate data from literature mining, expression analysis of dysregulated hsa_circ_0070269, circ-102,166, and miR-182 was performed in HCV induced HCC patients using RT-PCR.

Results: It was found that miR-182 was significantly upregulated and acts as an oncomiRNA in HCV induced HCC, and hsa_circ_0070269 and circ-102,166 were downregulated in HCV induced HCC. We have identified that miR-182 relative expression level was significantly high (p < 0.0029), while has_circ_0070269 (p < 0.002) and circ-102,166 (p < 0.002) were significantly downregulated in HCV-HCC patients as compared to expression in healthy individuals.

Conclusion: Our data revealed that miR-182 acts as an oncomiRNA in HCC development. Hsa_circ_0070269 and circ-102,166 are highly expressed in healthy controls compared to HCV induced HCC patients, can sponge miR-182 expression by acting as tumor suppressors, and can be used as biomarkers and targets for HCC treatment.

Keywords: HCV induced HCC; ceRNA; circRNA; hepatocellular carcinoma; miRNA; oncogenic pathways.

FIGURE 1

Flow chart of bioinformatic analysis.

FIGURE 2

Volcano plots of differentially expressed circRNA, miRNA and mRNA in HCC. (A) Dysregulated circRNA. (B) Dysregulated miRNA. (C) Dysregulated mRNA. Blue color indicates expression downregulation; red color indicates expression upregulation.

FIGURE 3

Structural pattern of circRNAs: (A) circ‐102,166; (B) hsa_circ‐0070269.

FIGURE 4

CircRNA‐miRNA‐mRNA network construction.

FIGURE 5

KEGG pathway analysis: (A) Bacterial invasion of epithelial cells; (B) Fc gamma R‐mediated phagocytosis; (C) choline metabolism in cancer; (D) parathyroid hormone synthesis, secretion, and action; and (F) Sphingolipid signaling pathway. The bar represents the −log10 (p‐value).

FIGURE 6

miRNAs and circRNAs role in pathogenesis. (A) miR‐182 role in different diseases by targeting different pathways. (B) miR‐182 role in HCC development by targeting tumor suppressor genes. (C) Hsa_circ_0070269 and miR‐182 in healthy liver. (D) Hsa_circ_0070269 and miR‐182 interaction HCC progression. (E) Circ‐102,166 and miR‐182 interaction in the healthy liver. (F) Circ‐102,166 and miR‐182 interaction in HCC progression.

FIGURE 7

Expression analysis of miR‐182, hsa_circ_0070269, and circ‐102,166. (A) MiR‐182 expression level in the blood samples of HCV‐HCC patients and healthy controls. (B) Hsa_circ_0070269 expression level in blood samples of HCV‐HCC patients and healthy individuals. (C) Circ‐102,166 expression level in blood samples of HCV‐HCC patients and healthy controls.

FIGURE 8

Comparison of the relative expression of the non‐coding RNA in HCV induced HCC and healthy controls. (A) Comparative relative expression of hsa_circ_0070269 and miR‐182. (B) Comparative relative expression of circ‐102,166 and miR‐182.