Expression analysis of liver-specific circulating microRNAs in HCV-induced hepatocellular Carcinoma in Egyptian patients

Figures & data

Table 1. Demographic and laboratory data of all patients and controls.

Variables	Controls (n = 40)	CHC (n = 42)	LC (n = 45)	HCC (n = 38)
Age	47.0 ± 3.5	46.4 ± 7.3	51.3 ± 4.7	48.1 ± 4.1
Gender (M:F)	3:1	3:2	7:5	6:4
Liver Function Tests (mean ± SD)
AST (U/L)	21.32 ± 1.07	46.14 ± 4.77 ^a	50.84 ± 5.00 ^a	76.40 ± 7.34 ^b
ALT (U/L)	23.16 ± 1.95	44.12 ± 5.50 ^a	50.84 ± 5.00 ^a	76.40 ± 7.34 ^b
ALP (U/L)	189 ± 41	331 ± 36 ^a	336 ± 48 ^a	420 ± 33 ^b
Albumin (g/dL)	4.4 ± 0.5	3.60 ± 0.74	3.8 ± 0.72	3.08 ± 0.48
Prothrombin Concentration	95.6 ± 3.4	89.6 ± 4.8	41.5 ± 11.1 ^b	69.4 ± 3.7 ^a
AFP (IU/mL)	3.12 ± 0.08	8.86 ± 0.11 ^a	10.11 ± 0.11 ^a	55.18 ± 0.44 ^b
DCP (mAU/ ml)	30.42 ± 0.70	121.49 ± 0.59 ^a	123.62 ± 0.38 ^a	456.52 ± 0.66 ^b

Data are expressed as mean ± standard deviations (SD).

Chronic hepatitis C (CHC); liver cirrhosis (LC); hepatocellular carcinoma (HCC).

Normal range for alanine aminotransferase (ALT) and aspartate aminotransferase (AST) is up to 40 IU/L.

Normal range for alkaline phosphatase (ALP) is up to 250 U/L.

Normal range for albumin is 3.5–5 g/dl.

Normal range for prothrombin concentration is 80–1 00%.

Normal range for alpha-fetoprotein (AFP) is 0.1–9.6 IU/mL.

mAU/ml = milli-absorbance unit/ml.

Normal range for des-γ-carboxy prothrombin (DCP) ≤40 mAU/ml.

^a p < 0.001 significant increase than control group.

^b p < 0.001 significant increase than CHC and LC groups.

Table 2. The qPCR expression levels of the serum microRNAs in the studied groups.

miRNAs	Normal	CHC	LC	HCC
miRNA-16	14.26 ± 0.69	24.09 ± 0.44^**	23.29 ± 0.46^**	22.35 ± 0.54^** ^a
miRNA-34a	27.32 ± 0.19	32.69 ± 0.34^**	30.01 ± 0.54^** ^b	32.50 ± 0.94^**
miRNA-125a	20.57 ± 0.54	96.01 ± 4.36^**	100.54 ± 0.81^**	29.96 ± 0.57^** ^d
miRNA-139	29.96 ± 0.57	94.63 ± 0.38^**	86.02 ± 0.40^** ^e	30.03 ± 0.43^d
miRNA-145	20.65 ± 0.52	85.31 ± 0.53^**	80.74 ± 0.59^**	20.64 ± 0.57^d
miRNA-199a	80.23 ± 0.72	330.38 ± 0.74^**	311.98 ± 0.72^**	66.16 ± 0.44^c d
miRNA-221	22.82 ± 0.38	27.17 ± 1.44^**	28.22 ± 0.41^**	28.51 ± 0.46^**

** p < 0.01 significant increase than control; ^ap < 0.01 significant decrease than CHC; ^bp < 0.01 significant decrease than CHC and HCC; ^cp < 0.01 significant decrease than control; ^dp < 0.01 significant decrease than CHC and LC; ^ep < 0.01 significant decrease than CHC

Figure 1. Box plot diagrams showing the expression of (A) miR-16, (B) miR-34a, (C) miR-125a, (D) miR-139, (E) miR-145, (F) miR-199a, and (G) miR-221 in Hepatitis C virus (HCV)-induced hepatocellular carcinoma (HCC) patients. The box indicates the 25^th and 75^th percentile of the data and the middle line indicates the median. A line extends from the minimum to the maximum value, excluding outliers that are displayed as separate points.

Table 3. Area Under the Curve (AUC), Confidence Interval (CI), and p-values for the circulating miRNAs.

MicroRNAs	AUC	CI 95%	p-value
miRNA-16	0.716 ± 0.05	0.604 – 0.829	<0.01
miRNA-34a	0.791 ± 0.04	0.695–0.888	<0.001
miRNA-125a	0.806 ± 0.04	0.729–0.884	<0.001
miRNA-139	0.863 ± 0.05	0.763–0.962	<0.001
miRNA-145	0.941 ± 0.02	0.902–0.980	<0.001
miRNA-199a	0.805 ± 0.03	0.704–0.906	<0.001
miRNA-221	0.633 ± 0.06	0.510–0.757	<0.05

Table 4. The sensitivity and specificity of the microRNAs.

MicroRNAs	Sensitivity	Specificity	PPV	NPV
miR-16	80.95	70.59	87.18	60.00
miR-34a	96.77	81.82	93.75	90.0
miR-125a	86.67	83.33	92.86	71.43
miR-139	85.71	64.29	82.76	69.23
miR-145	90.91	77.78	93.75	70.00
miR-199a	87.10	72.73	90.0	66.67
miR-221	90.32	81.82	93.33	75.0

PPV: Positive Predictive value

NPV: Negative Predictive Value