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Serum microRNA characterization identifies miR-885-5p as a potential marker for detecting liver pathologies.

Gui J, Tian Y, Wen X, Zhang W, Zhang P, Gao J, Run W, Tian L, Jia X, Gao Y - Clin. Sci. (2011)

Bottom Line: Five miRNAs (i.e. miR-885-5p, miR-574-3p, miR-224, miR-215 and miR-146a) that were up-regulated in the HCC and LC serum pools were selected and further quantified using real-time qPCR in patients with HCC, LC, CHB (chronic hepatitis B) or GC (gastric cancer) and in normal controls.The present study revealed that more than 110 miRNA species in the serum samples and wide distribution ranges of serum miRNAs were observed.No correlations between increased miR-885-5p and liver function parameters [AFP (α-fetoprotein), ALT (alanine aminotransferase), AST (aspartate aminotransferase) and GGT (γ-glutamyl transpeptidase)] were observed in patients with liver pathologies.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Biochemistry, Chinese PLA General Hospital, 28 Fuxing Rd, Beijing 100853, People's Republic of China.

ABSTRACT
Circulating miRNAs (microRNAs) are emerging as promising biomarkers for several pathological conditions, and the aim of this study was to investigate the feasibility of using serum miRNAs as biomarkers for liver pathologies. Real-time qPCR (quantitative PCR)-based TaqMan MicroRNA arrays were first employed to profile miRNAs in serum pools from patients with HCC (hepatocellular carcinoma) or LC (liver cirrhosis) and from healthy controls. Five miRNAs (i.e. miR-885-5p, miR-574-3p, miR-224, miR-215 and miR-146a) that were up-regulated in the HCC and LC serum pools were selected and further quantified using real-time qPCR in patients with HCC, LC, CHB (chronic hepatitis B) or GC (gastric cancer) and in normal controls. The present study revealed that more than 110 miRNA species in the serum samples and wide distribution ranges of serum miRNAs were observed. The levels of miR-885-5p were significantly higher in sera from patients with HCC, LC and CHB than in healthy controls or GC patients. miR-885-5p yielded an AUC [the area under the ROC (receiver operating characteristic) curve] of 0.904 [95% CI (confidence interval), 0.837-0.951, P<0.0001) with 90.53% sensitivity and 79.17% specificity in discriminating liver pathologies from healthy controls, using a cut off value of 1.06 (normalized). No correlations between increased miR-885-5p and liver function parameters [AFP (α-fetoprotein), ALT (alanine aminotransferase), AST (aspartate aminotransferase) and GGT (γ-glutamyl transpeptidase)] were observed in patients with liver pathologies. In summary, miR-885-5p is significantly elevated in the sera of patients with liver pathologies, and our data suggest that serum miRNAs could serve as novel complementary biomarkers for the detection and assessment of liver pathologies.

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Comparisons of levels of serum miRNAs in NC, HCC and LC groupsSerum abundances of miR-885-5p (A), miR-146a (B), miR-224 (C) and miR-574-3p (D) in an independent set of serum samples from NC (n=16), HCC (n=20) and LC (n=12) were quantified using real-time qPCR. Each qPCR was carried out in triplicate in 96-well plates. Expression levels of selected miRNAs were normalized to U6 snRNA and are presented as fold changes (2−ΔΔCt) above NC. Data are shown as the mean fold changes compared with the mean value in NC (±S.E.M.). A Mann–Whitney or Kruskal–Wallis test was used to determine statistical significance. ***P<0.0001, **P<0.001, *P<0.01.
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Figure 3: Comparisons of levels of serum miRNAs in NC, HCC and LC groupsSerum abundances of miR-885-5p (A), miR-146a (B), miR-224 (C) and miR-574-3p (D) in an independent set of serum samples from NC (n=16), HCC (n=20) and LC (n=12) were quantified using real-time qPCR. Each qPCR was carried out in triplicate in 96-well plates. Expression levels of selected miRNAs were normalized to U6 snRNA and are presented as fold changes (2−ΔΔCt) above NC. Data are shown as the mean fold changes compared with the mean value in NC (±S.E.M.). A Mann–Whitney or Kruskal–Wallis test was used to determine statistical significance. ***P<0.0001, **P<0.001, *P<0.01.

Mentions: Using U6 snRNA as the normalization control, we demonstrated that miR-885-5p was significantly higher in HCC and LC serum samples than in NC (P<0.0001), with a 6.5-fold increase in HCC and an 8.8-fold in LC (Figure 3A). In addition, increased amounts of serum miR-146a and miR-224 were observed in the HCC and LC groups (Figures 3B and 3C). However, no significant differences in the levels of miR-574-3p were observed among the NC, LC and HCC groups (Figure 3D), and the low level of miR-215 made it difficult to quantify its abundance in the sera accurately using real-time qPCR (most mean Ct values >35).


Serum microRNA characterization identifies miR-885-5p as a potential marker for detecting liver pathologies.

Gui J, Tian Y, Wen X, Zhang W, Zhang P, Gao J, Run W, Tian L, Jia X, Gao Y - Clin. Sci. (2011)

Comparisons of levels of serum miRNAs in NC, HCC and LC groupsSerum abundances of miR-885-5p (A), miR-146a (B), miR-224 (C) and miR-574-3p (D) in an independent set of serum samples from NC (n=16), HCC (n=20) and LC (n=12) were quantified using real-time qPCR. Each qPCR was carried out in triplicate in 96-well plates. Expression levels of selected miRNAs were normalized to U6 snRNA and are presented as fold changes (2−ΔΔCt) above NC. Data are shown as the mean fold changes compared with the mean value in NC (±S.E.M.). A Mann–Whitney or Kruskal–Wallis test was used to determine statistical significance. ***P<0.0001, **P<0.001, *P<0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2990200&req=5

Figure 3: Comparisons of levels of serum miRNAs in NC, HCC and LC groupsSerum abundances of miR-885-5p (A), miR-146a (B), miR-224 (C) and miR-574-3p (D) in an independent set of serum samples from NC (n=16), HCC (n=20) and LC (n=12) were quantified using real-time qPCR. Each qPCR was carried out in triplicate in 96-well plates. Expression levels of selected miRNAs were normalized to U6 snRNA and are presented as fold changes (2−ΔΔCt) above NC. Data are shown as the mean fold changes compared with the mean value in NC (±S.E.M.). A Mann–Whitney or Kruskal–Wallis test was used to determine statistical significance. ***P<0.0001, **P<0.001, *P<0.01.
Mentions: Using U6 snRNA as the normalization control, we demonstrated that miR-885-5p was significantly higher in HCC and LC serum samples than in NC (P<0.0001), with a 6.5-fold increase in HCC and an 8.8-fold in LC (Figure 3A). In addition, increased amounts of serum miR-146a and miR-224 were observed in the HCC and LC groups (Figures 3B and 3C). However, no significant differences in the levels of miR-574-3p were observed among the NC, LC and HCC groups (Figure 3D), and the low level of miR-215 made it difficult to quantify its abundance in the sera accurately using real-time qPCR (most mean Ct values >35).

Bottom Line: Five miRNAs (i.e. miR-885-5p, miR-574-3p, miR-224, miR-215 and miR-146a) that were up-regulated in the HCC and LC serum pools were selected and further quantified using real-time qPCR in patients with HCC, LC, CHB (chronic hepatitis B) or GC (gastric cancer) and in normal controls.The present study revealed that more than 110 miRNA species in the serum samples and wide distribution ranges of serum miRNAs were observed.No correlations between increased miR-885-5p and liver function parameters [AFP (α-fetoprotein), ALT (alanine aminotransferase), AST (aspartate aminotransferase) and GGT (γ-glutamyl transpeptidase)] were observed in patients with liver pathologies.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Biochemistry, Chinese PLA General Hospital, 28 Fuxing Rd, Beijing 100853, People's Republic of China.

ABSTRACT
Circulating miRNAs (microRNAs) are emerging as promising biomarkers for several pathological conditions, and the aim of this study was to investigate the feasibility of using serum miRNAs as biomarkers for liver pathologies. Real-time qPCR (quantitative PCR)-based TaqMan MicroRNA arrays were first employed to profile miRNAs in serum pools from patients with HCC (hepatocellular carcinoma) or LC (liver cirrhosis) and from healthy controls. Five miRNAs (i.e. miR-885-5p, miR-574-3p, miR-224, miR-215 and miR-146a) that were up-regulated in the HCC and LC serum pools were selected and further quantified using real-time qPCR in patients with HCC, LC, CHB (chronic hepatitis B) or GC (gastric cancer) and in normal controls. The present study revealed that more than 110 miRNA species in the serum samples and wide distribution ranges of serum miRNAs were observed. The levels of miR-885-5p were significantly higher in sera from patients with HCC, LC and CHB than in healthy controls or GC patients. miR-885-5p yielded an AUC [the area under the ROC (receiver operating characteristic) curve] of 0.904 [95% CI (confidence interval), 0.837-0.951, P<0.0001) with 90.53% sensitivity and 79.17% specificity in discriminating liver pathologies from healthy controls, using a cut off value of 1.06 (normalized). No correlations between increased miR-885-5p and liver function parameters [AFP (α-fetoprotein), ALT (alanine aminotransferase), AST (aspartate aminotransferase) and GGT (γ-glutamyl transpeptidase)] were observed in patients with liver pathologies. In summary, miR-885-5p is significantly elevated in the sera of patients with liver pathologies, and our data suggest that serum miRNAs could serve as novel complementary biomarkers for the detection and assessment of liver pathologies.

Show MeSH
Related in: MedlinePlus