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Journal of Molecular Medicine

, Volume 96, Issue 1, pp 85–96 | Cite as

Plasma circular RNA profiling of patients with gastric cancer and their droplet digital RT-PCR detection

  • Tianwen Li
  • Yongfu Shao
  • Liyun Fu
  • Yi Xie
  • Linwen Zhu
  • Weiliang Sun
  • Rui Yu
  • Bingxiu Xiao
  • Junming GuoEmail author
Original Article

Abstract

To observe the diagnostic values of circular RNAs (circRNAs), their expression profiles between gastric cancer patients’ plasma and healthy controls were first assessed by circRNA microarray. Then, circRNA levels were measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and RT-droplet digital PCR (RT-ddPCR), respectively. A total of 343 differentially expressed circRNAs were found. The top 10 elevated circRNAs in patients were hsa_circ_0088300, hsa_circ_0075825, hsa_circ_0019172, hsa_circ_0000220, hsa_circ_0035277, hsa_circ_0000301, hsa_circ_0000189, hsa_circ_0090080, hsa_circ_0001888, and hsa_circ_0001874. The top 10 reduced circRNAs were hsa_circ_0004771, hsa_circ_0001190, hsa_circ_0061276, hsa_circ_0092337, hsa_circ_0058495, hsa_circ_0061274, hsa_circ_0075829, hsa_circ_0080845, hsa_circ_0001006, and hsa_circ_0003764. In cancer and dysplasia tissues, hsa_circ_0001017 and hsa_circ_0061276 were downregulated. Their levels were significantly associated with distal metastasis. The area under receiver operating characteristic curve in combinative use was 0.966 with 95.5% sensitivity and 95.7% specificity. Patients with low plasma hsa_circ_0001017 or hsa_circ_0061276 had a much shorter overall survival than those with high levels. Patients whose plasma hsa_circ_0001017 or hsa_circ_0061276 levels recovered to normal after operation had a longer disease-free survival. Finally, the in vitro model indicated gastric cancer cells secreting circRNAs into plasma. In conclusion, RT-ddPCR is a potent non-invasive and absolute quantification method for simultaneous detection of multiple circRNAs. Hsa_circ_0001017 and hsa_circ_0061276 are new potential biomarkers for gastric cancer.

Key message

  • A total of 343 circRNAs are differentially expressed between gastric cancer patients’ plasma and healthy controls.

  • Hsa_circ_0001017 and hsa_circ_0061276 are downregulated in gastric cancer tissues.

  • The RT-ddPCR is a potent method for simultaneous detection of multiple circRNAs in plasma.

  • Hsa_circ_0001017 and hsa_circ_0061276 are potential biomarkers for gastric cancer.

Keywords

Biomarker  Gastric cancer  Non-invasive testing Digital polymerase chain reaction 

Notes

Acknowledgments

This study was supported by grants from the Applied Research Project on Nonprofit Technology of Zhejiang Province (no. 2016C33177), the Scientific Innovation Team Project of Ningbo (no. 2017C110019), National Natural Science Foundation of China (no. 81772279), and the K.C. Wong Magna Fund in Ningbo University.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

109_2017_1600_MOESM1_ESM.pdf (292 kb)
ESM 1 (PDF 291 kb)

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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular Biology, and Zhejiang Key Laboratory of Pathophysiology, Medical SchoolNingbo UniversityNingboChina
  2. 2.Department of Gastroenterology, the Affiliated Hospital of Medical School ofNingbo UniversityNingboChina
  3. 3.Department of Hepatology, Ningbo No. 2 Hospital and the Affiliated Hospital, Medical SchoolNingbo UniversityNingboChina
  4. 4.Ningbo Yinzhou People’s Hospital and the Affiliated Hospital, Medical SchoolNingbo UniversityNingboChina

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