Differential expression of circulating microRNAs in patients with cystic echinococcosis and screening for specific diagnostic biomarkers for the disease

Abstract

Abstract:

Objective To analyze the differential expression of circulating microRNAs (miRNAs) in patients with cystic echinococcosis (CE) and screen for potential diagnostic biomarkers for CE. Methods Serum miRNAs were extracted from 3 CE patients and 3 healthy controls (HCs) and analyzed with miRNA microarray using an Affymetrix miRNA4.0 chip to find miRNAs with differential expression. Results were verified on sera from 21 CE patients and 25 HCs using real-time PCR (qRT-PCR). Correlation between the miRNA and hydatid cyst weight or the number of daughter cysts was analyzed with Pearson’s correlation analysis. The IBM SPSS 19.0 software was used to generate the receiver operating characteristic curve (ROC) and analyze the area under curve (AUC). Results of microarrays revealed 172 miRNAs with an over 2-fold increase and 35 miRNAs with an over 50% decrease in CE sera as compared to the HC. qRT-PCR further quantitated the expression of hsa-miR-762, hsa-miR-3536, hsa-miR-6125, hsa-miR-6821-5P, hsa-miR-663a, hsa-miR-24-3P, hsa-miR-191-5P and hsa-miR-320a to be 10.28 ± 0.38, 5.11 ± 0.34, 11.02 ± 0.55, 11.87 ± 0.58, 5.31 ± 0.36, 5.75 ± 0.51, 5.75 ± 0.51 and 4.96 ± 0.36, respectively, all significantly higher than their HC counterparts (P < 0.05). ROC analysis showed that the AUC values of these 8 miRNA candidates were all above 8, implying their diagnostic potential for CE. Moreover, correlation analysis indicated that only miR-24-3p was positively correlated with both cyst weight (r = 0.5250, P < 0.05) and the number of daughter cysts (r = 0.6175, P < 0.05). Conclusion Eight circulating miRNAs have been identified as potential biomarkers for diagnosis of CE. MiR-24-3p may correlate with the progression and pathogenesis of CE.

Key words: Cystic echinococcosis, miRNA, Biomarker, MiR-24-3p

CLC Number:

R532.32

Meng-di REN, Yu-ye NING, Ming LI, Hao WANG, Wei ZHAO. Differential expression of circulating microRNAs in patients with cystic echinococcosis and screening for specific diagnostic biomarkers for the disease[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2017, 35(5): 423-428.

Figures/Tables 5

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引物名称 Primer name	序列 (5′→3′) Sequence (5′→3′)	长度/nt Length/nt
hsa-miR-762（S）	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACGCTCGG	50
hsa-miR-762（F）	TATTGAGGGGCTGGGGCC	18
hsa-miR-762（R）	CCAGTGCAGGGTCCGAGGTA	20
hsa-miR-3536（S）	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCCTCCC	50
hsa-miR-3536（F）	TATATCGGCGGGTGCGGG	18
hsa-miR-3536（R）	CCAGTGCAGGGTCCGAGGTA	20
hsa-miR-6125（S）	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACTCCGCC	50
hsa-miR-6125（F）	CACACAGCGGAAGGCGGA	18
hsa-miR-6125（R）	CCAGTGCAGGGTCCGAGGTA	20
hsa-miR-6821-5P（S）	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCCCCGC	50
hsa-miR-6821-5P（F）	TACACAGTGCGTGGTGGC	18
hsa-miR-6821-5P（R）	CCAGTGCAGGGTCCGAGGTA	20
hsa-miR-663a（S）	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACGCGGTC	50
hsa-miR-663a（F）	CTTCTAAGGCGGGGCGCC	18
hsa-miR-663a（R）	CCAGTCCAGGGTCCGAGGTA	20
hsa-miR-24-3P（S）	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCTGTTC	50
hsa-miR-24-3P（F）	CGCGCATGGCTCAGTTCA	18
hsa-miR-24-3P（R）	CCAGTGCAGGGTCCGAGGTA	20
hsa-miR-191-5P（S）	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCAGCTG	50
hsa-miR-191-5P（F）	CACAGTCAACGGAATCCC	18
hsa-miR-191-5P（R）	CCAGTGCAGGGTCCGAGGTA	20
hsa-miR-320a（S）	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACTCGCCC	50
hsa-miR-320a（F）	CGCAGCAAAAGCTGGGTT	18
hsa-miR-320a（R）	CCAGTGCAGGGTCCGAGGTA	20