Circulating miR-21-5p level has limited prognostic value in patients with hepatocellular carcinoma and is influenced by renal function

Table 1 Clinical and laboratory characteristics of hepatocellular carcinoma patients and hsa-miR-21-5p expression

		All patients	miR-21 low1	miR-21 high2	P value
Patient number:		91	47	44
Gender (n, %):	Women	17 (18.7%)	7 (14.9%)	10 (22.7%)	0.4231
	Men	74 (81.3%)	40 (85.1%)	34 (77.3%)
Age in yr	mean ± SD	67.91 ± 8.98	69.11 ± 8.84	66.64 ± 9.05	0.1912
Etiology (n, %):	Alcohol abuse	41 (45.1%)	22 (46.8%)	19 (43.2%)	0.7560
	Viral hepatitis	12 (13.2%)	5 (10.6%)	7 (15.9%)
	NASH	13 (14.3%)	8 (17.0%)	5 (11.4%)
	Rare or other cause3	25 (27.5%)	12 (25.5%)	13 (29.5%)
BCLC stage (n, %):	A	16 (17.6%)	7 (14.9%)	9 (20.5%)	0.7787
	B	37 (40.7%)	20 (42.6%)	17 (38.6%)
	C + D	38 (41.8%)	20 (42.6%)	18 (40.9%)
Child-Pugh score (n, %):	No liver cirrhosis	16 (17.6%)	8 (17.0%)	8 (18.2%)	0.7443
	A	45 (49.5%)	25 (53.2%)	20 (45.5%)
	B + C	30 (33.0%)	14 (29.8%)	16 (36.4%)
Treatment (n, %)4:	Therapy naïve	26 (28.6%)	11 (23.4%)	15 (34.1%)	0.3535
	Pretreated	65 (71.4%)	36 (76.6%)	29 (65.9%)
Ascites (n, %):	No ascites	58 (63.7%)	30 (63.8%)	28 (63.6%)	0.9999
	Ascites present	33 (36.3%)	17 (36.2%)	16 (36.4%)
Bilirubin (µmol/L):	mean ± SD	23.38 ± 31.73	24.27 ± 40.47	22.43 ± 18.78	0.7833
AFP (ng/mL):	mean ± SD	4593 ± 20162	540 ± 1533	8922 ± 28481	0.0469
INR5:	mean ± SD	1.076 ± 0.2127	1.034 ± 0.1450	1.122 ± 0.2611	0.0486
Platelets (Gpt/L):	mean ± SD	195.6 ± 118.7	182.7 ± 80.5	209.3 ± 149.0	0.2870
ALAT (μmol/Ls):	mean ± SD	0.7457 ± 0.4460	0.6955 ± 0.3425	0.7993 ± 0.5340	0.2697
ASAT (μmol/Ls):	mean ± SD	1.267 ± 1.041	1.040 ± 0.597	1.510 ± 1.330	0.0307
Hemoglobin (mmol/L):	mean ± SD	7.811 ± 1.254	7.655 ± 1.122	7.977 ± 1.375	0.2230
Creatinine (µmol/L):	mean ± SD	92.92 ± 67.95	102.3 ± 88.46	82.91 ± 33.08	0.1751
Albumin (g/L):	mean ± SD	36.29 ± 6.539	36.47 ± 5.668	36.10 ± 7.420	0.7938

¹miR-21 50% percentile.

²miR-21 > 50% percentile.

³Including hemochromatosis.

⁴Patients with different kind of pretreatments (for example hepatic resection, transarterial chemoembolization, selective internal radiation therapy and sorafenib) were included.

⁵For 30 patients we obtained no exact laboratory value of the International Normalized Ratio (only < 1.5). For these patients, we calculated the International Normalized Ratio based on the Quick. P values were calculated with unpaired t-test, Fisher’s exact test, and χ² test as appropriate. AFP: Alpha-fetoprotein; ALAT: Alanine aminotransferase; ASAT: Aspartate aminotransferase; BCLC: Barcelona Clinic Liver Cancer; INR: International Normalized Ratio; NASH: Non-alcoholic steatohepatitis; SD: Standard deviation.

Table 2 Correlation analyses of hsa-miR-21-5p and clinical or laboratory parameters

Parameter	2-ΔCt miR-21 / cel-miR-39
	Spearman r	Confidence interval	P value
Age in yr	-0.1147	-0.3188 to 0.09956	0.2789
Creatinine (µmol/L)	-0.2215	-0.4139 to -0.01013	0.0348
Bilirubin (µmol/L)	0.1484	-0.06547 to 0.3493	0.1603
ALAT (µmol/Ls)	0.1709	-0.0425 to 0.3694	0.1053
ASAT (µmol/Ls)	0.2854	0.07828 to 0.4689	0.0061
Albumin (g/L)	-0.01562	-0.2267 to 0.1969	0.8832
AFP (ng/mL)	0.1025	-0.1118 to 0.3077	0.3337
Hemoglobin (mmol/L)	0.1377	-0.07638 to 0.3396	0.1931
INR1	0.2065	-0.005615 to 0.4008	0.0496
Platelets (Gpt/L)	0.09185	-0.1224 to 0.2979	0.3865
2-ΔCt miR-122 / cel-miR-39	0.2624	0.05357 to 0.4493	0.0120

¹For 30 patients, we obtained no exact laboratory value of the International Normalized Ratio (only < 1.5). For these patients, we calculated the International Normalized Ratio based on the Quick. ALAT: Alanine aminotransferase; ASAT: Aspartate aminotransferase; AFP: Alpha-fetoprotein; INR: International Normalized Ratio; miR-21: Hsa-miR-21-5p.

Table 3 Comparison of studies with a statement about the prognostic value of blood hsa-miR-21-5p in patients with hepatocellular carcinoma

Characteristics	Qi et al[25]	Tomimaru et al[24]	Liu et al[32]	Wang et al[31]	Cho et al[33]	Zhang et al[18]	Current data
High miR-21 and prognosis	↔	Trend for worse prognosis	Worse prognosis	Worse prognosis	Better liver transplant-free survival	Worse prognosis	↔
Group allocation: Cut-off-value	NA	Cut-off based on ROC analyzes	50th percentile	50th percentile	50th percentile	NA	50th percentile
Origin of the cohort	Asia	Asia	Asia	Asia	Asia	Asia	Europe
Predominant etiology	HBV	HCV/HBV	HBV	HBV	HBV	NA	Mainly alcohol abuse
TNM reported	Yes	Yes		Yes	Yes	Yes
BCLC reported		Yes	Yes				Yes
Child-Pugh score reported	Yes	Yes	Yes		Yes		Yes
Sampling	Before surgery	Before surgery	Before TACE	No exact information, surgical resection	Before surgery or RFA	Pre- and postoperative serum samples	Therapy naïve and pretreated
Patients	70	126	136	97	120	46	91
Specimen	Serum	Plasma	Serum	Serum	Plasma	Serum	Serum
qPCR method	TaqMan®	TaqMan®	SYBRGreen	SYBR Green	TaqMan®	Unknown	TaqMan® SYBRGreen
Normalization	miR-16	miR-16	Quanto EC1, Quanto EC2	cel-miR-39	MiR-16	RNU6b	cel-miR-39
Publishing year	2011	2012	2014	2015	2017	2019	2020

BCLC: Barcelona Clinic Liver Cancer; HBV: Hepatitis B virus; HCC: Hepatocellular carcinoma; HCV: Hepatitis C virus; NA: Non-available; NASH: Non-alcoholic steatohepatitis; qPCR: Quantitative PCR; RFA: Radiofrequency ablation; ROC: Receiver operating characteristic; TACE: Transarterial chemoembolization; TNM: Tumor-Node-Metastasis Classification of Malignant Tumors.