TET2 controls chemoresistant slow-cycling cancer cell survival and tumor recurrence
Isabel Puig, … , Josep Tabernero, Héctor G. Palmer
Isabel Puig, … , Josep Tabernero, Héctor G. Palmer
Published August 31, 2018; First published June 26, 2018
Citation Information: J Clin Invest. 2018;128(9):3887-3905. https://doi.org/10.1172/JCI96393.
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Research Article Oncology

TET2 controls chemoresistant slow-cycling cancer cell survival and tumor recurrence

Abstract

Dormant or slow-cycling tumor cells can form a residual chemoresistant reservoir responsible for relapse in patients, years after curative surgery and adjuvant therapy. We have adapted the pulse-chase expression of H2BeGFP for labeling and isolating slow-cycling cancer cells (SCCCs). SCCCs showed cancer initiation potential and enhanced chemoresistance. Cells at this slow-cycling status presented a distinctive nongenetic and cell-autonomous gene expression profile shared across different tumor types. We identified TET2 epigenetic enzyme as a key factor controlling SCCC numbers, survival, and tumor recurrence. 5-Hydroxymethylcytosine (5hmC), generated by TET2 enzymatic activity, labeled the SCCC genome in carcinomas and was a predictive biomarker of relapse and survival in cancer patients. We have shown the enhanced chemoresistance of SCCCs and revealed 5hmC as a biomarker for their clinical identification and TET2 as a potential drug target for SCCC elimination that could extend patients’ survival.

Authors

Isabel Puig, Stephan P. Tenbaum, Irene Chicote, Oriol Arqués, Jordi Martínez-Quintanilla, Estefania Cuesta-Borrás, Lorena Ramírez, Pilar Gonzalo, Atenea Soto, Susana Aguilar, Cristina Eguizabal, Ginevra Caratù, Aleix Prat, Guillem Argilés, Stefania Landolfi, Oriol Casanovas, Violeta Serra, Alberto Villanueva, Alicia G. Arroyo, Luigi Terracciano, Paolo Nuciforo, Joan Seoane, Juan A. Recio, Ana Vivancos, Rodrigo Dienstmann, Josep Tabernero, Héctor G. Palmer

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Figure 5

SCCCs present enhanced chemoresistance.

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SCCCs present enhanced chemoresistance. (A–D) Chemoresistance evaluation...
(A–D) Chemoresistance evaluation of SCCCs and RCCCs in indicated models. (A–C) Analysis of apoptosis (A and C) and proportion of SCCCs (B and C) after chemotherapy exposure. OX, oxaliplatin; DTIC, light-activated dacarbazine; TMZ, temozolomide. Apoptosis measurements: SW1222 RCCC vehicle (VEH) vs. SCCC OX (P ≤ 0.01); RCCC OX vs. SCCC VEH (P ≤ 0.0001); MMLN9 RCCC DTIC vs. SCCC VEH (P ≤ 0.0001); e216 RCCC TMZ vs. SCCC VEH (P ≤ 0.0001). (D) Immunofluorescence of caspase-3 (CASP3) (n = 6 per group) treated or not treated with oxaliplatin. Arrowheads, SCCCs; asterisk, apoptotic areas. Scale bar: 100 μm. (E) qPCR of indicated genes. Data are represented as mean ± SD of triplicates. ND, not detected; r.u., relative units. (F) Apoptosis flow cytometric evaluation in RCCCs and SCCCs from CRC-SW1222-H2BeGFP cells growing as MTs. FTC, fumitremorgin C. Apoptosis measurements: RCCC VEH/FTC vs. RCCC OX/OX+FTC (P ≤ 0.0001); RCCC VEH/FTC vs. SCCC VEH/FTC (P ≤ 0.01); RCCC VEH/FTC vs. SCCC OX+FTC (P ≤ 0.001); RCCC OX/OX+FTC vs. SCCC VEH/FTC/OX/OX+FTC (P ≤ 0.0001); SCCC VEH/FTC vs. SCCC OX+FTC (P ≤ 0.0001); SCCC OX vs. SCCC OX+FTC (P ≤ 0.001). (G) Drug sensitivity of cancer cell lines according to PanC-SCCC signature scores. Adjusted Wilcoxon test. (H) Disease-free survival of chemo-treated high-risk stage II/III colon cancer patients (GSE39582, n = 151) according to CRC-SCCC signature score. HR, hazard ratio. Cox proportional hazards model. (A–C and F) Data are represented as mean ± SEM. (A, B, E, and F) Data were obtained from triplicates of 3 independent experiments. (A–C, E, and F) Blue bars, RCCCs; green bars, SCCCs. (A and F) 1-way ANOVA. (B, C, and E) 2-tailed Student’s t test. (A–C and E–G) *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001.