LncRNAs and <em>EGFRvIII</em> sequestered in TEPs enable blood-based NSCLC diagnosis

Abstract

BackgroundTissue biopsy-based cancer diagnosis has limitations because of the fact that tumor tissues are in constant evolution and extremely heterogeneous. The current study was aimed to examine whether tumor-educated blood platelets (TEPs) might be a potential all-in-one source for blood-based cancer diagnostics to overcome the limitations of conventional cancer biopsy.MethodsIn the present study, we evaluated the expression pattern of MAGI2 antisense RNA 3 (MAGI2-AS3) and ZNFX1 antisense RNA 1 (ZFAS1) in both plasma and platelets of 101 non-small-cell lung cancer (NSCLC) patients. Receiver operating characteristic (ROC) curve was generated to evaluate their diagnostic potential. In addition, epidermal growth factor receptor (EGFR) mutations were detected in DNA and RNA samples of platelets for companion diagnostics.ResultsOur results showed that the levels of MAGI2-AS3 and ZFAS1 in both plasma and platelets of NSCLC patients were significantly downregulated than those in healthy controls. A positive correlation of long noncoding RNA expression was observed between platelets and plasma (r=0.738 for MAGI2-AS3, r=0.751 for ZFAS1, respectively). By ROC analysis, we found that molecular interrogation of MAGI2-AS3 and ZFAS1 in TEPs and plasma can offer valuable diagnostic performance for NSCLC patients (area under the ROC curve [AUC]MAGI2-AS3= 0.853/0.892, and AUCZFAS1=0.780/0.744 for diagnosing adenocarcinoma and squamous cell carcinoma cases from controls, respectively). Clinicopathologic characteristic analysis further revealed that MAGI2-AS3 level significantly correlated with tumor–node–metastasis (TNM) stage (p=0.001 in TEPs, p=0.003 in plasma), lymph-node metastasis (p=0.016 in TEPs, p=0.023 in plasma), and distant metastasis (p=0.045 in TEPs, p=0.045 in plasma), while ZFAS1 level was only correlated with TNM stage (p=0.005 in TEPs, p=0.044 in plasma). Furthermore, EGFRvIII RNA existed in both TEPs and plasma, but EGFR intracellular mutations cannot be detected in DNA of TEPs isolated from NSCLC.ConclusionOur data suggested that TEP is a promising source for NSCLC diagnosis and companion diagnostics.

<p>Long Noncoding MAGI2-AS3 Suppresses Several Cellular Processes of Lung Squamous Cell Carcinoma Cells by Regulating miR-374a/b-5p/CADM2 Axis</p>

“…15,26 The downregulation of MAGI2-AS3 has been proved in NSCLC including LUAD and LUSC. 16,17 A recent research validated the tumor-suppressive role of MAGI2-AS3 in NSCLC based on LUAD cell lines. 18 However, the role of MAGI2-AS3 in LUSC is yet to be shown.…”

“…[13][14][15] Importantly, previous studies have identified that MAGI2-AS3 is down-regulated in NSCLC samples, including LUAD and LUSC samples. 16,17 Moreover, we identified through GEPIA online tool based on TCGA data that MAGI2-AS3 was downregulated in LUSC samples versus normal samples. These findings indicated that MAGI2-AS3 might participate in LUSC.…”

Post-Transcriptional Expression Control in Platelet Biogenesis and Function

“…In NSCLC, the expression of lncRNAs was analyzed in platelets in order to identify novel biomarkers of lung cancer. A selection of five lncRNAs was identified and further measurements revealed a differential expression of two lncRNAs, namely, Membrane-Associated Guanylate Kinase Inverted 2-Antisense RNA 3 ( MAGI2-AS3 ) and Zinc Finger NFX1-Type Containing 1 Antisense RNA 1 ( ZFAS1 ), in platelets of NSCLC patients compared to healthy subjects [ 94 ]. Both lncRNAs have been assigned tumor suppressive functions in human breast cancer [ 95 , 96 , 97 ], whereas ZFAS1 seems to be a potent oncogenic lncRNA in other tumor types [ 98 ].…”

“…In platelets and plasma of NSCLC patients, MAGI2-AS3 and ZFAS1 were downregulated and their expression significantly correlated with tumor stage. In addition, MAGI2-AS3 expression significantly correlated with lymph-node and distant metastasis and the authors suggested that both platelet-derived lncRNAs could be used as potential diagnostic biomarkers in NSCLC [ 94 ]. However, the molecular function of these and other lncRNAs in platelets is currently not well understood and it is tempting to speculate about a potential transfer of lncRNAs from platelets to diverse other cell-types to modulate expression programs and phenotypes.…”

Δ133p53/FLp53 predicts poor clinical outcome in esophageal squamous cell carcinoma

“…Total RNA was extracted using an RNAprep Pure FFPE Kit (TIANGEN Biotech, Beijing, China) according to the manufacturer's instructions. Total RNA of 300 µL serum was isolated by using a Liquid Total RNA Isolation Kit (BioTeke, Beijing, China) as we described previously [27] and treated with DNase I prior to reverse transcription. The concentration of RNA was measured by Nanodrop 2000 spectrophotometer (Thermo Fisher) following puri cation with RNeasy mini kit (QIAGEN).…”