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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.

Tong YS, Wang XW, Zhou XL, Liu ZH, Yang TX, Shi WH, Xie HW, Lv J, Wu QQ, Cao XF - Mol. Cancer (2015)

Bottom Line: 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.Furthermore, plasma levels of POU3F3, HNF1A-AS1 and SPRY4-IT1 were significantly higher in ESCC patients compared with normal controls.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%).

View Article: PubMed Central - PubMed

Affiliation: Department of Surgical Oncology, Nanjing Frist Hospital, Nanjing Medical University, Nanjing, Jiangsu, China. tongyusuo@163.com.

ABSTRACT

Background: 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.

Methods: 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.

Results: 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%).

Conclusions: 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.

No MeSH data available.


Related in: MedlinePlus

Origin of circulating lncRNAs. (A) Quantitative real time polymerase chain reaction (qPCR) was used to measure ESCC-related lncRNAs expression in five esophageal cells. GAPDH was used as a normalization control. (B) ESCC-related lncRNAs could be secreted into the cell culture medium. Data presented as relative lncRNAs fold change. (C) ESCC-related lncRNAs could enter into the circulation of xenograft-bearing mice. (D) Spearman’s rank correlation scatter plot of ESCC-related lncRNAs levels in tumor samples and plasma. Data were presented as △Ct values normalized to GAPDH. (E) ESCC-related lncRNAs expressions were significantly declined in post-operative samples compared with that in pre-operative samples. (F) Effect of delayed blood centrifugation on plasma ESCC-related lncRNAs expressions. At room temperature, ESCC-related lncRNAs levels of unfiltered plasma was slightly increased from 0 h to 6 h, but then significantly decreased at 24 h when compared to 6 h. *p < 0.05.
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Fig5: Origin of circulating lncRNAs. (A) Quantitative real time polymerase chain reaction (qPCR) was used to measure ESCC-related lncRNAs expression in five esophageal cells. GAPDH was used as a normalization control. (B) ESCC-related lncRNAs could be secreted into the cell culture medium. Data presented as relative lncRNAs fold change. (C) ESCC-related lncRNAs could enter into the circulation of xenograft-bearing mice. (D) Spearman’s rank correlation scatter plot of ESCC-related lncRNAs levels in tumor samples and plasma. Data were presented as △Ct values normalized to GAPDH. (E) ESCC-related lncRNAs expressions were significantly declined in post-operative samples compared with that in pre-operative samples. (F) Effect of delayed blood centrifugation on plasma ESCC-related lncRNAs expressions. At room temperature, ESCC-related lncRNAs levels of unfiltered plasma was slightly increased from 0 h to 6 h, but then significantly decreased at 24 h when compared to 6 h. *p < 0.05.

Mentions: In the first experiment, qPCR was used to measure the POU3F3, HNF1A-AS1 and SPRY4-IT1 expression in different five esophageal cell lines (including four ESCC cells: KYSE30, KYSE70, KYSE450, and Eca 109, and one normal human esophageal epithelial cell line: HET-1A), and each cell culture medium which was incubated for 1, 2, and 3d (Figure 5A and B). Data presented in Figure 5B indicated that ESCC-related lncRNAs could enter into the cell culture medium at a detectable level and were steadily increased among the three incubation time points in the all four ESCC cell lines; however, no significant changes were observed in normal esophageal epithelial cell culture medium.


Identification of the long non-coding RNA POU3F3 in plasma as a novel biomarker for diagnosis of esophageal squamous cell carcinoma.

Tong YS, Wang XW, Zhou XL, Liu ZH, Yang TX, Shi WH, Xie HW, Lv J, Wu QQ, Cao XF - Mol. Cancer (2015)

Origin of circulating lncRNAs. (A) Quantitative real time polymerase chain reaction (qPCR) was used to measure ESCC-related lncRNAs expression in five esophageal cells. GAPDH was used as a normalization control. (B) ESCC-related lncRNAs could be secreted into the cell culture medium. Data presented as relative lncRNAs fold change. (C) ESCC-related lncRNAs could enter into the circulation of xenograft-bearing mice. (D) Spearman’s rank correlation scatter plot of ESCC-related lncRNAs levels in tumor samples and plasma. Data were presented as △Ct values normalized to GAPDH. (E) ESCC-related lncRNAs expressions were significantly declined in post-operative samples compared with that in pre-operative samples. (F) Effect of delayed blood centrifugation on plasma ESCC-related lncRNAs expressions. At room temperature, ESCC-related lncRNAs levels of unfiltered plasma was slightly increased from 0 h to 6 h, but then significantly decreased at 24 h when compared to 6 h. *p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4631113&req=5

Fig5: Origin of circulating lncRNAs. (A) Quantitative real time polymerase chain reaction (qPCR) was used to measure ESCC-related lncRNAs expression in five esophageal cells. GAPDH was used as a normalization control. (B) ESCC-related lncRNAs could be secreted into the cell culture medium. Data presented as relative lncRNAs fold change. (C) ESCC-related lncRNAs could enter into the circulation of xenograft-bearing mice. (D) Spearman’s rank correlation scatter plot of ESCC-related lncRNAs levels in tumor samples and plasma. Data were presented as △Ct values normalized to GAPDH. (E) ESCC-related lncRNAs expressions were significantly declined in post-operative samples compared with that in pre-operative samples. (F) Effect of delayed blood centrifugation on plasma ESCC-related lncRNAs expressions. At room temperature, ESCC-related lncRNAs levels of unfiltered plasma was slightly increased from 0 h to 6 h, but then significantly decreased at 24 h when compared to 6 h. *p < 0.05.
Mentions: In the first experiment, qPCR was used to measure the POU3F3, HNF1A-AS1 and SPRY4-IT1 expression in different five esophageal cell lines (including four ESCC cells: KYSE30, KYSE70, KYSE450, and Eca 109, and one normal human esophageal epithelial cell line: HET-1A), and each cell culture medium which was incubated for 1, 2, and 3d (Figure 5A and B). Data presented in Figure 5B indicated that ESCC-related lncRNAs could enter into the cell culture medium at a detectable level and were steadily increased among the three incubation time points in the all four ESCC cell lines; however, no significant changes were observed in normal esophageal epithelial cell culture medium.

Bottom Line: 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.Furthermore, plasma levels of POU3F3, HNF1A-AS1 and SPRY4-IT1 were significantly higher in ESCC patients compared with normal controls.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%).

View Article: PubMed Central - PubMed

Affiliation: Department of Surgical Oncology, Nanjing Frist Hospital, Nanjing Medical University, Nanjing, Jiangsu, China. tongyusuo@163.com.

ABSTRACT

Background: 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.

Methods: 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.

Results: 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%).

Conclusions: 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.

No MeSH data available.


Related in: MedlinePlus