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. 2022 Feb 11;21:46. doi: 10.1186/s12943-022-01511-1

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Identification and characterization of circRHOBTB3 in CRC. (A) Heatmap of differentially expressed circRNAs in sera from CRC, HCC, and PAAD patients compared with healthy donors. (B) Venn diagram of significantly upregulated circRNAs in CRC, HCC, and PAAD serum samples. (C) Expression of hsa_circ_000744 and its host gene in serum samples from CRC, HCC, PAAD patients and healthy donors. (D) Sketch map and primers for circRHOBTB3. (E) Sanger sequencing for confirmation of the specific back-splicing site of circRHOBTB3. (F) RNase R treatment, RT-PCR and RT-qPCR for confirmation of circRHOBTB3. (G) Detection of circRHOBTB3 in serum samples from CRC patients by RT-qPCR (levels normalized to those of GAPDH). (H) Detection of circRHOBTB3 and RHOBTB3 in paired normal and CRC tissue in the verification cohort by RT-qPCR (levels normalized to those of GAPDH). (I) Kaplan-Meier survival analysis of CRC patients according to the expression level of circRHOBTB3 using the log-rank test (n = 69). All experiments were repeated for three times, data were shown as mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001, NS P > 0.05, in Student’s test (C, F, G), or paired Student’s test (H)