Research Paper Volume 13, Issue 12 pp 16178—16197

GPR30-mediated HMGB1 upregulation in CAFs induces autophagy and tamoxifen resistance in ERα-positive breast cancer cells

Phosphorylated Akt is involved in HMGB1-mediated autophagy. (A) The Beclin1, p62, and LC3, autophagy-related marker proteins in MCF-7 cells cultured with different conditioned medium (CM) from CAFs, (B) with recombinant HMGB1, or with (C) HMGB1 neutralizing antibody detected by western blotting. (Control: MCF-7 cells cultured with DMEM phenol red-free medium; CAF-shNC (CM): MCF-7 cells cultured with CM from CAF-shNC cells; G1+CAF-sh NC (CM): MCF-7 cells cultured with CM from CAF-shNC cells treated with G1; CAF-shGPR30 (CM): MCF-7 cells cultured with CM from CAF-shGPR30 cells; G1+CAF-sh GPR30 (CM): MCF-7 cells cultured with CM from CAF-shGPR30 cells treated with G1. *P D) Phosphorylated Akt was detected with the addition of LY294002 by western blot analysis. All experiments were independently repeated at least 3 times.

Figure 4. Phosphorylated Akt is involved in HMGB1-mediated autophagy. (A) The Beclin1, p62, and LC3, autophagy-related marker proteins in MCF-7 cells cultured with different conditioned medium (CM) from CAFs, (B) with recombinant HMGB1, or with (C) HMGB1 neutralizing antibody detected by western blotting. (Control: MCF-7 cells cultured with DMEM phenol red-free medium; CAF-shNC (CM): MCF-7 cells cultured with CM from CAF-shNC cells; G1+CAF-sh NC (CM): MCF-7 cells cultured with CM from CAF-shNC cells treated with G1; CAF-shGPR30 (CM): MCF-7 cells cultured with CM from CAF-shGPR30 cells; G1+CAF-sh GPR30 (CM): MCF-7 cells cultured with CM from CAF-shGPR30 cells treated with G1. *P < 0.05, vs CAF(CM) and CAF-shNC (CM **P < 0.01, vs CAF(CM) and CAF-shNC (CM); ∆P < 0.01, G1+CAF-shNC (CM) (n = 3). (D) Phosphorylated Akt was detected with the addition of LY294002 by western blot analysis. All experiments were independently repeated at least 3 times.