The online version of this article (doi:10.1186/1476-4598-14-3) contains supplementary material, which is available to authorized users.
The authors declare that they have no conflict of interests.
X.C. and Y.T. conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript, X.Z. conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing; X.W., Z.L., T.Y., and W.S. data analysis and interpretation; J.L., H.X. and Q.W. provision of study materials. All authors read and approved the final manuscript.
Recent studies have demonstrated that long non-coding RNAs (lncRNAs) were present in the blood of cancer patients and have shown great potential as powerful and non-invasive tumor markers. However, little is known about the value of lncRNAs in the diagnosis of esophageal squamous cell carcinoma (ESCC). We hypothesized that ESCC-related lncRNAs might be released into the circulation during tumor initiation and could be utilized to detect and monitor ESCC.
Ten lncRNAs (HOTAIR, AFAP1-AS1, POU3F3, HNF1A-AS1, 91H, PlncRNA1, SPRY4-IT1, ENST00000435885.1, XLOC_013104 and ENST00000547963.1) which previously found to be differently expressed in esophageal cancer were selected as candidate targets for subsequent circulating lncRNA assay. A four-stage exploratory study was conducted to test the hypothesis: (1) optimization of detected method to accurately and reproducibly measure ESCC-related lncRNAs in plasma and serum; (2) evaluation of the stability of circulating lncRNAs in human plasma or serum; (3) exploration the origin of ESCC-related lncRNAs in vitro and in vivo; (4) evaluation the diagnostic power of circulating lncRNAs for ESCC.
ESCC-related lncRNAs were detectable and stable in plasma of cancer patients, and derived largely from ESCC tumor cells. Furthermore, plasma levels of POU3F3, HNF1A-AS1 and SPRY4-IT1 were significantly higher in ESCC patients compared with normal controls. By receiver operating characteristic curve (ROC) analysis, among the three lncRNAs investigated, plasma POU3F3 provided the highest diagnostic performance for detection of ESCC (the area under the ROC curve (AUC), 0.842; p < 0.001; sensitivity, 72.8%; specificity, 89.4%). Moreover, use of POU3F3 and SCCA in combination could provide a more effective diagnosis performance (AUC, 0.926, p < 0.001, sensitivity, 85.7%; specificity, 81.4%). Most importantly, this combination was effective to detect ESCC at an early stage (80.8%).
Plasma POU3F3 could serve as a potential biomarker for diagnosis of ESCC, and the combination of POU3F3 and SCCA was more efficient for ESCC detection, in particular for early tumor screening.
Additional file 1: Table S1: Raw Ct values of 21 pairs of ESCC tumor tissues and adjacent normal tissues. (DOC 32 KB)12943_2014_1498_MOESM1_ESM.doc
Additional file 2: Table S2: Correlation between GAPDH level (raw Ct value) in human plasma and clinicopathological factors of normal controls and ESCC patients. (DOC 32 KB)12943_2014_1498_MOESM2_ESM.doc
Additional file 3: Text S1: Supplementary data. (DOCX 16 KB)12943_2014_1498_MOESM3_ESM.docx
Additional file 4: Figure S1: Sequencing Results of Plasma qPCR Products of POU3F3 (A), HNF1A-AS1 (B) and SPRY4-IT1 (C). (TIFF 3 MB)12943_2014_1498_MOESM4_ESM.tiff
Additional file 5: Figure S2: Comparison plasma level of ENST00000435885 and AFAP1-AS1 between ESCC and normal control groups. ENST00000435885 (A) and AFAP1-AS1 (B) expression showed no significant differences between ESCC and normal controls. Data presented as △Ct values normalized to GAPDH. (TIFF 0 bytes)12943_2014_1498_MOESM5_ESM.tiff
Additional file 6: Figure S3: Effect of delayed processing of blood on ESCC-related lncRNAs expression. ●, unfiltered plasma incubated at 4°C, △, filtered plasma incubated at room temperature. The symbols represented the means at specified time points. (TIFF 3 MB)12943_2014_1498_MOESM6_ESM.tiff
Additional file 7: Table S3: Correlation between ESCC-related lncRNAs expression (△Ct) in plasma and clinicopathological characteristics of 143 ESCC patients. (DOC 64 KB)12943_2014_1498_MOESM7_ESM.doc
Additional file 8: Table S4: Clinical characteristics of study population. (DOC 54 KB)12943_2014_1498_MOESM8_ESM.doc
Additional file 9: Table S5: Checklist MIQE. (DOC 116 KB)12943_2014_1498_MOESM9_ESM.doc
Additional file 10: Table S7: qPCR target information. (DOC 54 KB)12943_2014_1498_MOESM10_ESM.doc
Additional file 11: Table S6: Primers sequences. (DOC 116 KB)12943_2014_1498_MOESM11_ESM.doc
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- Identification of the long non-coding RNA POU3F3 in plasma as a novel biomarker for diagnosis of esophageal squamous cell carcinoma
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