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. 2021 Dec 6;75(6):1386–1401. doi: 10.1002/hep.32177

FIGURE 5.

FIGURE 5

mTORC1‐mediated translation of HIF‐1α and SREBP1 drives IR‐R in HCC cells. (A) Synergy of rapamycin (2 μM, 48 h) with IR in radioresistant cells by apoptosis measurements. (B,C) Restorative effect of M‐HCl (10 μM) (B) or CRLS1 overexpression (C) on clonogenic survival of radioresistant cell lines treated with rapamycin (1 μM) and exposed to the indicated dose of IR. Survival data were normalized to respective unirradiated controls. (D) MG132 (20 μM, 24 h) or chloroquine (20 μM, 24 h) treatment on indicated cells with or without 8 Gy IR. (E) The ratio of HIF‐1α and SREBP1 mRNA loaded on polysomes to their total mRNA levels by quantitative RT‐PCR analysis in IR or IR plus rapamycin–treated conditions (2 μM, 24 h). Each value was normalized to tubulin loaded on the polysome and total tubulin expression. (F) Protein levels of HIF‐1α and SREBP1 in radioresistant cells with 4E‐BP1 or S6K knocked down by siRNA with 8 Gy IR. Data are represented as mean ± SEM of at least three replicates. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Abbreviations: Con, control; CQ, chloroquine; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; (M), mature; ns, not significant; P, parental; (P), precursor; Rapa, rapamycin [Color figure can be viewed at wileyonlinelibrary.com]