| Objective: The purpose of this study is to investigate the correlation between the acoustic features of papillary thyroid carcinoma and miRNA-144-5p level,and to analyze the potential molecular regulation mechanism and clinical value of miRNA-144-5p in thyroid carcinoma.Methods:1.Correlation between ultrasound characteristics and miRNA-144-5p expression.Two-dimensional ultrasound image characteristics of the oven(number,size,edge,aspect ratio,echogenicity,calcification,etc.)and contrast-enhanced ultrasound features(enhanced mode,enhanced level,the increase of time,etc.)of 24 cases of papillary thyroid cancer were retrospectively summarized.The expression levels of miRNA-144-5p in 24 cases of PTC and 19 non-cancerous tissues were detected by rt-q PCR,and the expression level of miRNA-144-5p in the two groups was compared by independent sample t test.Fisher's exactprobability method was used to analyze the correlation between the ultrasound features and the expression of miRNA-144-5p.2.acquired and analyzed the date from The Cancer Genome Atlas(TCGA),Gene Expression Omnibus(GEO)and Arrayexpress database.The sequencing data of miRNA-144-5p and clinical parameters of patients were obtained from TCGA databases.The chip data were obtained from the GEO and Arrayexpress databases.The expression differences of miRNA-144-5p between TC and non-cancer tissues were analyzed by independent sample t test.The correlation between the expression level of miRNA-144-5p and clinicopathological parameters was assessed by chi-square test.The Kaplan-Meier curve was used to analyze difference of the overall survival rate between the miRNA-144-5p high expression group and the miRNA-144-5p low expression group.Standard mean difference(SMD)and summary receiver operating characteristic curve(SROC)were used for meta-analysis of the above data.Then,the target genes of miRNA-144-5p were predicted by 12 online prediction platforms.Gene ontology(GO)was used to analyze the function of miRNA-144-5p,and the prospective mechanism of miRNA-144-5p was analyzed by the Kyoto encyclopedia of genes and genomes(KEGG)pathway.The gene express in the most enriched pathway of KEGG pathway was analyzed by TCGA date,and the Over-expression mRNA was defined as the target gene.Pearson's correlation coefficient was used to test the correlation between miRNA-144-5p and target genes.The Human Protein Altas was used to verify the protein expression of the target genes in thyroid carcinoma.Results:The two-dimensional ultrasound images of 25 cases of control group lesions characterized by mixed echo(64%),regular edge(84%),A/T < 1(100%),the CEUS images of 25 cases of control group lesions characterized by Partial equienhancement and Partial non-enhancement(80%),clear border(100%)and heterogeneous enhancement(80%).The two-dimensional ultrasound images of 24 cases of PTC lesions characterized by irregular edges(83.3%),A/T?1(37.5%),solid or very low echo(16.7%),with microcalcification(58.3%),the CEUS images of 24 cases of PTC lesions characterized by hypoenhancement(70.8%),uneven enhancement(75%),unclear strengthen boundary(54.1%)?The result of RT-qPCR showed that the miRNA-144-5p expression was remarkably lower in TC(P< 0.01),which was consistent with the TCGA data.ultrasound features A/T ? 1 and hypoenhancement were associated with miRNA-144-5p lower expression(P = 0.045 and 0.034,respectively).Combined GEO and Arrayexpress database chip data,compared with non-cancer tissues,miRNA-144-5p expression was remarkably lower in the following three chips compared with non-cancer tissues : GSE40807,GSE73182 and e-mtab-736(P< 0.001).TCGA data analysis showed that the expression of miRNA-144-5p was significantly lower in TC than in non-cancer tissues(P<0.001).The area under the ROC curve was 0.922(P< 0.001),and the difference was statistically significant.The clinical parameter analysis showed that the low expression of miRNA-144-5p was related to the advanced stage(III and IV stage)of TNM stage(P=0.005).However,other clinical parameters such as age,gender,number of lesions,distant metastasis,lymph node metastasis had no significant correlation(P = 0.378,0.378,0.378,0.545 and 0.838,respectively).With the combination of GEO,Arrayexpress and TCGA data,the meta-analysis showed that the total sensitivity of miRNA-144-5p was 0.90(95%CI:0.86-0).91),total specificity 0.88(95%CI:0.81-0.93),total positive likelihood ratio(PLR)5.97(95%CI: 3.70-9.63),total negative likelihood ratio(NLR)0.16(95%CI: 0.07-0.36),and the total diagnostic ratio(DOR)44.52(95%CI:The SMD =-1.786(95%CI:-2.379-(-1.192),P<0.001,random effect model),and the area under the SROC curve was 0.94(95%CI 0.95-0.98 P<0.01),indicating that miRNA-144-5p was used to distinguish between thyroid cancer and non-cancer.In addition,through 12 different predicting tools,the genes that overlaped over 9 tools were enrolled.Finally,283 possible target genes were obtained.Subsequently,KEGG analysis showed that the target genes were mainly enriched in ‘Proteoglycans in cancer' pathway,which contained 8genes(CAV2,ANK2,TFAP4,PPP1R12 A,FZD4,ITPR1,FZD6,FN1).These above 8 genes were tested by TCGA expression analysis,and only FN1 was highly expressed in thyroid cancer,which could be the target gene.Pearson's correlation coefficient test results suggested that miRNA-144-5p was significantly negatively correlated with FN1(r =-0.363;95% CI: 0.432-(0.289)),P < 0.001).Conclusion: Ultrasound image characteristics of PTC A/T ? 1 and hypoenhancement were correlated with the lower expression of miRNA-144-5p,are risk factors for preoperative risk assessment of PTC.The results of large data analysis shows that miRNA-144-5p was low in PTC,and it may be possible for miRNA-144-5p to participate in the occurrence and development of TC through FN1,but more experiments are needed to prove this. |
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