Plasma circular RNA profiling of patients with gastric cancer and their droplet digital RT-PCR detection
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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.
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.
KeywordsBiomarker Gastric cancer Non-invasive testing Digital polymerase chain reaction
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.
- 3.Allemani C, Weir HK, Carreira H, Harewood R, Spika D, Wang XS, Bannon F, Ahn JV, Johnson CJ, Bonaventure A et al (2015) Global surveillance of cancer survival 1995-2009: analysis of individual data for 25, 676, 887 patients from 279 population-based registries in 67 countries (CONCORD-2). Lancet 385:977–1010CrossRefPubMedGoogle Scholar
- 14.Shen F, Sun B, Kreutz JE, Davydova EK, Du W, Reddy PL, Joseph LJ, Ismagilov RF (2011) Multiplexed quantification of nucleic acids with large dynamic range using multivolume digital RT-PCR on a rotational SlipChip tested with HIV and hepatitis C viral load. J Am Chem Soc 133:17705–17712CrossRefPubMedPubMedCentralGoogle Scholar
- 22.Janku F, Huang HJ, Fujii T, Shelton DN, Madwani K, Fu S, Tsimberidou AM, Piha-Paul SA, Wheler JJ, Zinner RG et al (2017) Multiplex KRASG12/G13 mutation testing of unamplified cell-free DNA from the plasma of patients with advanced cancers using droplet digital polymerase chain reaction. Ann Oncol 28:642–650CrossRefPubMedGoogle Scholar
- 26.Capello M, Bantis LE, Scelo G, Zhao Y, Li P, Dhillon DS, Patel NJ, Kundnani DL, Wang H, Abbruzzese JL et al (2017) Sequential validation of blood-based protein biomarker candidates for early-stage pancreatic cancer. J Natl Cancer Inst 109. https://doi.org/10.1093/jnci/djw266
- 27.Min QH, Chen XM, Zou YQ, Zhang J, Li J, Wang Y, Li SQ, Gao QF, Sun F, Liu J et al (2017) Differential expression of urinary exosomal microRNAs in IgA nephropathy. J Clin Lab Anal. https://doi.org/10.1002/jcla.22226
- 28.Raitoharju E, Seppala I, Lyytikainen LP, Viikari J, Ala-Korpela M, Soininen P, Kangas AJ, Waldenberger M, Klopp N, Illig T et al (2016) Blood hsa-miR-122-5p and hsa-miR-885-5p levels associate with fatty liver and related lipoprotein metabolism—the young Finns study. Sci Rep 6:38262CrossRefPubMedPubMedCentralGoogle Scholar