Abstract
This study evaluated differences in the clinical appearance of patients with hepatocellular carcinoma (HCC) based on plasma level and regulation of microRNAs (miRNA-29c, miRNA-21, and miRNA-155). The observational-analytical study with a cross-sectional design was conducted on 36 HCC patients and 36 healthy controls. The blood samples were collected from 2 Province Hospitals (Dr. Sardjito Hospital and Prof. Dr. Margono Soekarjo Hospital) for HCC and the Blood Bank Donor of the Indonesian Red Cross for 36 healthy controls. These blood samples were treated as follows: plasma isolation, RNA isolation, cDNA synthesis, quantification by qRT-PCR using a sequence-specific forward primer, and normalization of miRNA using housekeeping-stably miRNA-16. There were only 27 HCC patients with complete clinical variables (neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), platelet count, albumin, C-reactive protein (CRP), and cholinesterase (ChE)) that were able to analyses for regulation miRNAs based on its fold change expression miRNA target. All 27 HCC subjects were follow-up until 3-years of monitoring for their overall survival. The miRNA plasma expression was analyzed by Bio-Rad CFX 96 Manager software to determine the cycle of quantification, followed by the calculation of expression levels using Livak's methods. Data were analyzed using STATA 11.0, with a significant value of p<0.05. The miRNAs expression of HCC subjects were lower than that healthy control subjects in miRNA-29c (down-regulation 1.83-fold), higher than that healthy control subjects in miRNA 21 and miRNA-155 (up-regulation, 1.74-fold; 1.55-fold) respectively. NLR, CRP, ChE, and platelet count showed a significant difference in miRNA-29c regulation, though neutrophil count showed a significant difference in miRNA-21 and miRNA-155 regulation (p<0.05). Conclusion: Plasma biomarkers: miRNA-21 and miRNA-155 might be potential biomarkers as onco-miR in HCC subjects, while miRNA-29c might act as a tumor suppressor. Significant evidence was identified with clinical progression based on the regulation of miRNAs, which was consistent with miRNA -29c.
Highlights
Hepatocellular carcinoma (HCC) is the fifth most common of the major malignant liver tumors in the world and the third leading cause of mortality-related cancer
All 36 HCC subjects who enrolled in the study continued to examine the miRNAs expression levels (ΔmiRNA) for miRNA 29c-3p, miRNA 21-5p, and miRNA 155-5p [11, 14]
The ΔCq miRNA 155-5p and ΔCq miRNA 21-5p in HCC patients were lower than healthy controls, but only ΔCq miRNA 155-5p showed significantly different values with p
Summary
Hepatocellular carcinoma (HCC) is the fifth most common of the major malignant liver tumors in the world and the third leading cause of mortality-related cancer. Plasma levels of miRNA-122 and miRNA-21 have strong correlations with the degree of fibrosis in HCV genotype-4 patients They can be considered as a potential biomarker for the early detection of Plasma miRNA-29c, miRNA-21 and miRNA-155 in clinical progression of Hepatocellular Carcinoma patients hepatic fibrosis. The serum level of miRNA-21 is able for detecting the liver necroinflammatory activity event there was not the presence of HCC [12]. Activity and synthesis of ChE would be reduced if there is any cellular liver dysfunction following with damaging of the cell membrane and this is a cause of diminished reserve function. A previous study revealed that the alteration of ChE in the cirrhotic liver may reflect the changes in the pathophysiology of synthesis at the protein and miRNA levels, even though enzymatic activity does not change [21]. A combination of inflammation biomarkers (CRP and NLR) can be used as a predictor of tumor response and survival [28]
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