通过 WNT2 表达区分结直肠腺瘤和增生性息肉。
Distinguishing colorectal adenoma from hyperplastic polyp by WNT2 expression.
机构信息
Department of Digestive Diseases, Huashan Hospital, Fudan University, Shanghai, China.
State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China.
出版信息
J Clin Lab Anal. 2021 Oct;35(10):e23961. doi: 10.1002/jcla.23961. Epub 2021 Sep 3.
BACKGROUND
Colorectal adenoma (CRA) is a classical premalignant lesion, with high incidence and mainly coexisting with hyperplastic polyp (HPP). Hence, this study aimed to distinguish CRA from HPP by molecular expression profiling and advance the prevention of CRA and its malignance.
METHODS
CRA and paired HPP biopsies were collected by endoscopy. Through RNA-sequencing (RNA-seq), the differentially expressed genes (DEGs) were obtained. Functional enrichment analysis was performed based on the DEGs. The STRING database and Cytoscape were used to construct the protein-protein interaction (PPI) network and perform module analysis. Hub genes were validated by real-time quantitative PCR (RT-qPCR) and immunohistochemistry. The ROC curve was drawn to establish the specificity of the hub genes.
RESULTS
485 significant DEGs were identified including 133 up-regulated and 352 down-regulated. The top 10 up-regulated genes were DLX5, MMP10, TAC1, ACAN, TAS2R38, WNT2, PHYHIPL, DKK4, DUSP27, and ABCA12. The top 10 down-regulated genes were SFRP2, CHRDL1, KBTBD12, RERGL, DPP10, CLCA4, GREM2, TMIGD1, FEV, and OTOP3. Wnt signaling pathway and extracellular matrix (ECM) were up-regulated in CRA. Three hub genes including WNT2, WNT5A, and SFRP1 were filtered out via Cytoscape. Further RT-qPCR and immunohistochemistry confirmed that WNT2 was highly expressed in CRA. The area under the ROC curve (AUC) at 0.98 indicated the expression level of WNT2 as a candidate to differ CRA from HPP.
CONCLUSION
Our study suggests Wnt signaling pathway and ECM are enriched in CRA, and WNT2 may be used as a novel biomarker for distinguishing CRA from HPP and preventing the malignance of CRA.
背景
结直肠腺瘤（CRA）是一种经典的癌前病变，发病率高，主要与增生性息肉（HPP）共存。因此，本研究旨在通过分子表达谱分析来区分 CRA 和 HPP，从而推进 CRA 及其恶性的预防。
方法
通过内镜收集 CRA 和配对的 HPP 活检。通过 RNA 测序（RNA-seq）获得差异表达基因（DEGs）。基于 DEGs 进行功能富集分析。使用 STRING 数据库和 Cytoscape 构建蛋白质-蛋白质相互作用（PPI）网络并进行模块分析。通过实时定量 PCR（RT-qPCR）和免疫组织化学验证核心基因。绘制 ROC 曲线以建立核心基因的特异性。
结果
鉴定出 485 个显著的 DEGs，包括 133 个上调基因和 352 个下调基因。上调基因的前 10 位分别为 DLX5、MMP10、TAC1、ACAN、TAS2R38、WNT2、PHYHIPL、DKK4、DUSP27 和 ABCA12。下调基因的前 10 位分别为 SFRP2、CHRDL1、KBTBD12、RERGL、DPP10、CLCA4、GREM2、TMIGD1、FEV 和 OTOP3。CRA 中 Wnt 信号通路和细胞外基质（ECM）上调。通过 Cytoscape 筛选出三个核心基因，包括 WNT2、WNT5A 和 SFRP1。进一步的 RT-qPCR 和免疫组织化学证实 WNT2 在 CRA 中高表达。ROC 曲线下面积（AUC）为 0.98，表明 WNT2 的表达水平可作为区分 CRA 和 HPP 的候选标志物。
结论
本研究表明 Wnt 信号通路和 ECM 在 CRA 中富集，WNT2 可作为区分 CRA 和 HPP 并预防 CRA 恶性转化的新型生物标志物。
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