PDF(Pubmed)
Abstract：
OBJECTIVE: The aim of our study is to find a better way to identify a group of papillary thyroid carcinoma (PTC) with more aggressive behaviors and to provide a prediction model for lymph node metastasis to assist in clinic practice.
METHODS: Targeted sequencing of DNA/RNA was used to detect genetic alterations. Gene expression level was measured by quantitative real-time PCR, western blotting or immunohistochemistry. CCK8, transwell assay and flow cytometry were used to investigate the effects of concomitant gene alterations in PTC. LASSO-logistics regression algorithm was used to construct a nomogram model integrating radiomic features, mutated genes and clinical characteristics.
RESULTS: 172 high-risk variants and 7 fusion types were detected. The mutation frequencies in BRAF, TERT, RET, ATM and GGT1 were significantly higher in cancer tissues than benign nodules. Gene fusions were detected in 16 samples (2 at the DNA level and 14 at the RNA level). ATM mutation (ATMMUT) was frequently accompanied by BRAFMUT, TERTMUT or gene fusions. ATMMUT alone or ATM co-mutations were significantly positively correlated with lymph node metastasis. Accordingly, ATM knock-down PTC cells bearing BRAFV600E, KRASG12R or CCDC6-RET had higher proliferative ability and more aggressive potency than cells without ATM knock-down in vitro. Furthermore, combining gene alterations and clinical features significantly improved the predictive efficacy for lymph node metastasis of radiomic features, from 71.5 to 87.0%.
CONCLUSIONS: Targeted sequencing of comprehensive genetic alterations in PTC has high prognostic value. These alterations, in combination with clinical and radiomic features, may aid in predicting invasive PTC with higher accuracy.
METHODS: Targeted sequencing of DNA/RNA was used to detect genetic alterations. Gene expression level was measured by quantitative real-time PCR, western blotting or immunohistochemistry. CCK8, transwell assay and flow cytometry were used to investigate the effects of concomitant gene alterations in PTC. LASSO-logistics regression algorithm was used to construct a nomogram model integrating radiomic features, mutated genes and clinical characteristics.
RESULTS: 172 high-risk variants and 7 fusion types were detected. The mutation frequencies in BRAF, TERT, RET, ATM and GGT1 were significantly higher in cancer tissues than benign nodules. Gene fusions were detected in 16 samples (2 at the DNA level and 14 at the RNA level). ATM mutation (ATMMUT) was frequently accompanied by BRAFMUT, TERTMUT or gene fusions. ATMMUT alone or ATM co-mutations were significantly positively correlated with lymph node metastasis. Accordingly, ATM knock-down PTC cells bearing BRAFV600E, KRASG12R or CCDC6-RET had higher proliferative ability and more aggressive potency than cells without ATM knock-down in vitro. Furthermore, combining gene alterations and clinical features significantly improved the predictive efficacy for lymph node metastasis of radiomic features, from 71.5 to 87.0%.
CONCLUSIONS: Targeted sequencing of comprehensive genetic alterations in PTC has high prognostic value. These alterations, in combination with clinical and radiomic features, may aid in predicting invasive PTC with higher accuracy.
摘要：
目的：我们研究的目的是寻找一种更好的方法来识别一组具有更积极行为的甲状腺乳头状癌（PTC），并提供淋巴结转移的预测模型，以帮助临床实践。
方法：DNA/RNA的靶向测序用于检测遗传改变。通过实时定量PCR检测基因表达水平,蛋白质印迹或免疫组织化学。使用CCK8，transwell测定和流式细胞术研究PTC中伴随基因改变的影响。采用LASSO-logistics回归算法构建了一个整合放射学特征的列线图模型,突变基因和临床特征。
结果：共检测到172种高危变异和7种融合类型。BRAF中的突变频率,TERT,RET,ATM和GGT1在癌组织中明显高于良性结节。在16个样品中检测到基因融合(DNA水平2个,RNA水平14个)。ATM突变（ATMMUT）经常伴有BRAFMUT，TERTMUT或基因融合。单独ATMMUT或ATM共突变与淋巴结转移呈显著正相关。因此，带有BRAFV600E的ATM击倒PTC电池，KRASG12R或CCDC6-RET在体外比没有ATM敲低的细胞具有更高的增殖能力和更积极的效力。此外，结合基因改变和临床特征显着提高了放射学特征对淋巴结转移的预测功效，从71.5到87.0%。
结论：PTC综合基因改变的靶向测序具有较高的预后价值。这些改动,结合临床和影像学特征，可以帮助以更高的准确性预测侵入性PTC。
方法：DNA/RNA的靶向测序用于检测遗传改变。通过实时定量PCR检测基因表达水平,蛋白质印迹或免疫组织化学。使用CCK8，transwell测定和流式细胞术研究PTC中伴随基因改变的影响。采用LASSO-logistics回归算法构建了一个整合放射学特征的列线图模型,突变基因和临床特征。
结果：共检测到172种高危变异和7种融合类型。BRAF中的突变频率,TERT,RET,ATM和GGT1在癌组织中明显高于良性结节。在16个样品中检测到基因融合(DNA水平2个,RNA水平14个)。ATM突变（ATMMUT）经常伴有BRAFMUT，TERTMUT或基因融合。单独ATMMUT或ATM共突变与淋巴结转移呈显著正相关。因此，带有BRAFV600E的ATM击倒PTC电池，KRASG12R或CCDC6-RET在体外比没有ATM敲低的细胞具有更高的增殖能力和更积极的效力。此外，结合基因改变和临床特征显着提高了放射学特征对淋巴结转移的预测功效，从71.5到87.0%。
结论：PTC综合基因改变的靶向测序具有较高的预后价值。这些改动,结合临床和影像学特征，可以帮助以更高的准确性预测侵入性PTC。
