Identification Of NONHSAT076754 In Papillary Thyroid Cancer Lymph Node Metastasis And Its Role In Aiding Ultrasound Diagnosis

ObjectivesThyroid cancer is a common endocrine malignancy.Its global incidence has increased a lot because of the development of ultrasonography and the application of ultrasoundguided fine needle aspiration(US-FNA),and most of the new cases are thyroid microcarcinoma.The increasement of the thyroid cancer cases causes a larger volume of thyroid surgery,while the motality remains the same.Lymph node metastasis(LNM)is an indicator of recurrence in papillary thyroid cancer(PTC).An efficient evaluation of LNM in PTC would make a real difference to the surgical management as well as the postoperative surveillance of PTC patients.Ultrasound is the most important method in examing thyroid nodules,however,its sensitivity in evaluation of LNM is relatively low.It will be of great value to fine a new way to increase the sensitivity of Ultrasound in diagnosing LNM and also improve the effect of treatment.Long noncoding RNA(lnc RNA)plays a critical role in tumorigenesis and metastasis,which was confirmed in various cancers.The clinical significance of lnc RNAs as biomarker was also affirmed.The present study aims to investigate the role of lnc RNA in LNM of PTC,and also identify the value of US-FNAlnc RNA in diagosing LNM of PTC.MethodsLnc RNAs expression profile of PTC with LNM was acquired by using whole transcriptome sequencing.Bioinformatic analysis was conducted to predict the target gene of lnc RNA as well as lnc RNA-m RNA coepxression network.The expression levels of candidate lnc RNAs were verified in PTC patients with LNM and without LNM using realtime PCR and fluorescence in situ hybridization(FISH).In vitro,the expression of lnc RNA was knocked down or overexpressed and then the affect on biological behaviors such as cell migration,proliferation etc.were observed;the expression of predicted target gene FN1(FN1)was also detected.In addition,the role of FN1 and TWSG1 on PTC metastasis was investigated.Moreover,the value of ultrasonography in predicting LNM of PTC was calculated.The study collected a large volume of FNA samples of PTC and the expressions of lnc RNA,FN1,TWSG1 were compared between LNM group and non LNM group of PTC.Furthermore,the correlations of clinicopathological characteristics of these patients with lnc RNA,FN1,TWSG1 expression level were analyzed.The correlation study of ultrasonographical characteristics with lnc RNA was also conducted.After that,the value of US-FNA-lnc RNA in evaluation of LNM of PTC before surgery was explored.The clinical significance of FN1,TWSG1 as diagnostic biomarkers of LNM of PTC were also demonstrated.ResultsThis study acquired lnc RNAs expression profile based on 18 cases of PTC specimens.Bioinformatic analysis showed that FN1 was the cis gene of NONHSAT076754,which was a hub in the lnc RNA-m RNA coexpression network.In the verification study,NONHSAT076754,FN1 and TWSG1 was found to be highly expressed in PTC patients with LNM compared to those without LNM.FISH analysis also showed that NONHSAT076754 located in cytoplasm.In vitro study indicated that overexpression of NONHSAT076754,FN1 and TWSG1 promoted PTC proliferation,invasion and migration.TWSG1 also promoted the expression of tumor vascular factor and matrix degrading enzyme.Moreover,analysis of ultrasonographic images from 458 PTC cases showed that the diagnostic value of US in LNM of PTC was only 65.72%.Correlation study showed that high expressions of NONHSAT076754,FN1 and TWSG1 were correlated with PTC LNM.Multivariate logistic regression analysis showed that NONHSAT076754 was an independent risk factor of LNM of PTC.In addition,the value of US-FNANONHSAT076754 in the preoperative diagnosis of PTC LNM was 87.5%,and the clinical value of FN1 and TWSG1 as tumor biomarker in predicting LNM of PTC before surgery were 81.1% and 78% respectively.ConclusionNONHSAT076754,FN1 and TWSG1 are overexpressed in PTC with LNM compared to PTC without LNM.The overexpression of NONHSAT076754,FN1 and TWSG1 promote PTC proliferation and metastasis.NONHSAT076754 regulates PTC metastasis through positively regulating FN1 and TWSG1.NONHSAT076754 is a potential therapeutic target of PTC LNM.The overexpression of NONHSAT076754,FN1 and TWSG1 correlate with LNM of PTC.NONHSAT076754 improves the diagnostic accuracy of ultrasonography in predicting LNM of PTC before surgery.FN1 and TWSG1 are also potential diagnostic biomarkers of PTC with LNM.