Fig. 7: TRMT6/TRMT61A complex inhibitor thiram suppresses self-renewal of liver CSCs and tumor growth.
a Quantitative LC-MS/MS analysis of m1A/A in total RNA and tRNA purified from PLC/PRF/5 cells treated with vehicle or drugs at IC50. Statistical results as means ± SD, n = 5 biologically independent samples. Exact P values from left to right: 0.0048, 0.0086, 0.0037, 0.0090. b Oncosphere forming assays of HCC primary cells following thiram or vehicle treatment. Scale bar, 100 μm. Right panel represents statistical results as means ± SD. n = 15 independent experiments. c Representative magnetic resonance imaging (MRI) images of independent experimental cohorts, of day 0 and day 15 of mice enrolled in treatment with vehicle (DMSO), thiram (1.6 mg kg−1), GSK3787 (5 mg kg−1) or thiram combined with GSK3787. Red circles and asterisk indicate visible tumor area that was used to calculate tumor volumes. Scale bar, 1 cm. Tumor volumes were calculated on the basis of MRI images from mice bearing liver tumor with a matched initial tumor volume (Right panel). Graph shows means ± SD. n = 6 mice per group. Exact P values from left to right: 0.00065, 0.028. d Tumor volumes were calculated from mice treated with vehicle or atorvastatin (Atorva., 10 mg kg−1). Graph shows means ± SD. n = 6 mice per group. Exact P value: 0.0073. e Survival curve derived from mice bearing HCC PDC tumors, treated with vehicle (n = 11), thiram (n = 12), GSK3787 (n = 10), or thiram combined with GSK3787 (n = 12). P values for Kaplan–Meier curves were determined using a two-sided log-rank test. Exact P values from up to down: 0.00011, 0.025, 0.018. f Survival curve derived from mice bearing HCC PDC tumors, treated with vehicle (n = 10) and atorvastatin (n = 10). P value for Kaplan–Meier curves were determined using a two-sided log-rank test. Exact P value: 0.011. g, i Representative immunohistochemical images of m1A and PPARδ levels in PDC orthotopic engraft mouse models treated with vehicle, thiram, GSK3787, thiram combined with GSK3787 (combination) or atorvastatin (Atorva.). Scale bar, 100 μm. h, j Graphs of numbers of m1A positive (m1A+) cells per tumor nodule after treatment. Data are means ± SD. n = 20. Exact P values from left to right: <0.0001, 0.13, 0.85 (h); and 0.58 (j). *P < 0.05; **P < 0.01; ***P < 0.001; NS not significant (P > 0.05) by two-tailed Student’s t-test.
