Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Sep 18;47(9):e184.
doi: 10.1038/emm.2015.68.

Serum exosomal microRNAs as novel biomarkers for hepatocellular carcinoma

Won Sohn  1 , Jonghwa Kim  1 , So Hee Kang  1 , Se Ra Yang  1 , Ju-Yeon Cho  1 , Hyun Chin Cho  2 , Sang Goon Shim  2 , Yong-Han Paik  1   3
Affiliations

Serum exosomal microRNAs as novel biomarkers for hepatocellular carcinoma

Won Sohn et al. Exp Mol Med. .

Abstract

Recent studies have shown that circulating microRNAs are a potential biomarker in various types of malignancies. The aim of this study was to investigate the feasibility of using serum exosomal microRNAs as novel serological biomarkers for hepatocellular carcinoma (HCC) in patients with chronic hepatitis B (CHB). We measured the serum exosomal microRNAs and serum circulating microRNAs in patients with CHB (n=20), liver cirrhosis (LC) (n=20) and HCC (n=20). Serum exosomal microRNA was extracted from 500 μl of serum using an Exosome RNA Isolation kit. The expression levels of microRNAs were quantified by real-time PCR. The expression levels of selected microRNAs were normalized to Caenorhabditis elegans microRNA (Cel-miR-39). The serum levels of exosomal miR-18a, miR-221, miR-222 and miR-224 were significantly higher in patients with HCC than those with CHB or LC (P<0.05). Further, the serum levels of exosomal miR-101, miR-106b, miR-122 and miR-195 were lower in patients with HCC than in patients with CHB (P=0.014, P<0.001, P<0.001 and P<0.001, respectively). There was no significant difference in the levels of miR-21 and miR-93 among the three groups. Additionally, the serum levels of circulating microRNAs showed a smaller difference between HCC and either CHB or LC. This study suggests that serum exosomal microRNAs may be used as novel serological biomarkers for HCC.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Expression of CD63, CD9 and calnexin by western blotting analysis. Lane 1: isolated exosomal pellets from the serum of healthy humans (positive control), lane 2: isolated exosomal pellets from the serum of patients, and lanes 3 and 4: Huh-7 cell lysates.
Figure 2
Figure 2
Serum exosomal microRNAs from patients with chronic hepatitis B (CHB), liver cirrhosis (LC) and hepatocellular carcinoma (HCC). The levels of serum exosomal microRNA were measured by real-time quantitative PCR. The values of the relative gene expression for target microRNA were normalized to Cel-miR-39 and calculated using the 2−ΔΔCT method. *The level of serum exosomal microRNA was increased in the HCC group compared with the CHB group (P<0.05). **The level of serum exosomal microRNA was decreased in the HCC group compared with the CHB group (P<0.05). †The level of serum exosomal microRNA was increased in the HCC group compared with the LC group (P<0.05). The values are expressed as the mean±s.e.m.
Figure 3
Figure 3
The distribution of upregulated exosomal microRNAs (miR-18a, -221, -222 and -224) in patients with CHB, LC and HCC. Each serum exosomal microRNA was compared between two groups (CHB vs HCC, CHB vs LC and HCC vs LC). The levels of serum exosomal microRNA were measured by real-time quantitative PCR. The values of relative gene expression for target microRNA were normalized to Cel-miR-39 and calculated using the 2−ΔΔCT method. P<0.05 was considered statistically significant. CHB, chronic hepatitis B; LC, liver cirrhosis; HCC, hepatocellular carcinoma.
Figure 4
Figure 4
The distribution of downregulated exosomal microRNAs (miR-101, -106b, -122 and -195) in patients with CHB, LC and HCC. Each serum exosomal microRNA was compared between two groups (CHB vs HCC, CHB vs LC and HCC vs LC). The levels of serum exosomal microRNA were measured by real-time quantitative PCR. The values of relative gene expression for target microRNA were normalized to Cel-miR-39 and calculated using the 2−ΔΔCT method. P<0.05 was considered statistically significant. CHB, chronic hepatitis B; LC, liver cirrhosis; HCC, hepatocellular carcinoma.
Figure 5
Figure 5
Serum circulating microRNAs in chronic hepatitis B (CHB), liver cirrhosis (LC) and hepatocellular carcinoma (HCC). The levels of serum circulating microRNA were measured by real-time quantitative PCR. The values of the relative gene expression for target microRNA were normalized to Cel-miR-39 and calculated using the 2−ΔΔCT method. *The level of serum microRNA was increased in the LC group compared with the CHB group (P<0.05). There was no significant difference in the level of serum circulating microRNA between the CHB and HCC groups. The value was expressed as the mean±s.e.m. The correlation of microRNA expression between serum exosomal and circulating microRNAs was investigated in each group. †A high correlation coefficient was observed in miR-21 in CHB (r=0.636, P=0.048), miR-221 in LC (r=0.770, P=0.009), miR-222 in HCC (r=0.508, P=0.022) and miR-224 in HCC (r=0.547, P=0.012).
See this image and copyright information in PMC

References

    1. 1Bushati N, Cohen SM. microRNA functions. Annu Rev Cell Dev Biol 2007; 23: 175–205. - PubMed
    1. 2Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004; 116: 281–297. - PubMed
    1. 3Benhamou Y, Di Martino V, Bochet M, Colombet G, Thibault V, Liou A et al. Factors affecting liver fibrosis in human immunodeficiency virus-and hepatitis C virus-coinfected patients: impact of protease inhibitor therapy. Hepatology 2001; 34: 283–287. - PubMed
    1. 4Ruan K, Fang X, Ouyang G. MicroRNAs: novel regulators in the hallmarks of human cancer. Cancer Lett 2009; 285: 116–126. - PubMed
    1. 5Nana-Sinkam SP, Croce CM. Clinical applications for microRNAs in cancer. Clin Pharmacol Ther 2013; 93: 98–104. - PubMed