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. 2021 Jun 6;20:86. doi: 10.1186/s12943-021-01370-2

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© The Author(s) 2021

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 — FHIT is highly methylated in ATL but not TSP patient samples and correlates with decreased FHIT expression. a Red and black arrows represent the amplicon for BGS and MSPCR, respectively of individual CpG islands (blocks). A representation of MSPCR for HDs (n = 10), ACs (n = 10), TSP/HAM (n = 10), and ATL (n = 10) samples are shown. Disease type is marked in columns A-D; while individual patients are marked in rows A-J. MSPCR amplifying the unmethylated (U) or methylated (M) FHIT gene product are shown; individual patients are noted by brackets. b Representation of BGS of HDs, ACs, TSP, and ATL patients (ATL-A: acute ATL, ATL-C, chronic ATL, and ATL-S, smoldering ATL). U vs M alleles are noted by white and black boxes, respectively. Horizontal rows represent sequencing of a single colony. c FHIT methylation in ATL (n = 198), ATL subtypes, TSP (n = 136), AC (n = 89), and HD (n = 39) from MSPCR. d Statistical analysis of FHIT methylation in HD, AC, and ATL samples. p-values are determined by Chi-Square test. e Representation of the overall percentage of M CpG islands (n = 32) in FHIT for HD (n = 12), AC (n = 10), TSP (n = 21), and ATL samples (chronic n = 46, acute n = 25). Results represent the average percentage of CpG islands in the analyzed portion of FHIT from BGS. The ANOVA f-ratio value is 16.67206, with a p-value of < .00001. Individual post hoc Tukey’s HSD p-values are noted when statistically significant between groups. f Distinct regions of the FHIT gene are methylated in ATL samples. The results are scored from the percentage of FHIT gene methylation at an individual CpG island from an average of approximately 20 acute ATL patients. g Quantitative real-time PCR performed on HDs (n = 38) and mostly acute ATL samples (n = 67) for FHIT gene expression. h Quantitative real-time PCR for FHIT gene expression in HDs (n = 15), methylated ATL (ATL-M; n = 16), and TSP/HAM (TSP; n = 14) samples. i Quantitative real-time PCR for FHIT gene expression in unmethylated (n = 7) vs methylated (n = 20) ATL patients. For (g-i), results were normalized to GAPDH expression and represent a fold change from a HD considered as “1”. p-values were calculated using a two-tailed, unequal variance T-test. FHIT CpG status was determined by MSPCR