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

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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.

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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.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, and Zhejiang Key Laboratory of Pathophysiology, Medical School, Ningbo University, Ningbo, 315211, China

    Tianwen Li, Yi Xie, Linwen Zhu, Rui Yu, Bingxiu Xiao & Junming Guo

  2. Department of Gastroenterology, the Affiliated Hospital of Medical School of, Ningbo University, Ningbo, 315020, China

    Yongfu Shao

  3. Department of Hepatology, Ningbo No. 2 Hospital and the Affiliated Hospital, Medical School, Ningbo University, Ningbo, 315010, China

    Liyun Fu

  4. Ningbo Yinzhou People’s Hospital and the Affiliated Hospital, Medical School, Ningbo University, Ningbo, 315040, China

    Weiliang Sun

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  1. Tianwen Li
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Correspondence to Junming Guo.

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Li, T., Shao, Y., Fu, L. et al. Plasma circular RNA profiling of patients with gastric cancer and their droplet digital RT-PCR detection. J Mol Med 96, 85–96 (2018). https://doi.org/10.1007/s00109-017-1600-y

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  • DOI: https://doi.org/10.1007/s00109-017-1600-y

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