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. 2021 Jun 16;11(12):3966–3982. doi: 10.1016/j.apsb.2021.06.007

Figure 7.

Figure 7

Inhibition of mitophagy enhances the anticancer efficacy of magnolol. (A) SH-SY5Y cells were transiently transfected with Scr siRNA and mixed PINK1 plus Parkin siRNA for 48 h, and then treated with DMSO and magnolol 100 μmol/L for 24 h. Cell pellets were subsequently collected, and cell death was quantified using PI live exclusion staining. Data are presented as mean ± SD (n = 3). ∗∗P < 0.01 (two-way ANOVA). (B) SH-SY5Y cells treated with indicated treatments [DMSO, magnolol 100 μmol/L, wortmannin (wort) 5 μmol/L, vacuolin-1 1 μmol/L, Baf-A1 100 nmol/L, CQ 20 μmol/L, and combined agents] for 24 h were collected and cell death was quantified using PI live exclusion staining. Data are presented as mean ± SD (n = 3). ∗∗∗P < 0.001 (one-way ANOVA). (C) Representative photos of SH-SY5Y xenografted tumors. Mice were intraperitoneally treated with saline (Ctrl), magnolol (35 mg/kg), wortmannin (0.35 mg/kg), magnolol + wortmannin for 21 days. Scale bar: 5 mm. (D) Tumor volumes were measured every three days following the treatment in (B). Values are represented as mean ± SD from at least 5 tumors in each group. ∗∗P < 0.01, ∗∗∗P < 0.001 (two-way ANOVA). (E) Tumor weights were analyzed after 21 days' treatment in (B). Values are represented as mean ± SD from at least 5 tumors in each group. ∗∗P < 0.01, ∗∗∗P < 0.001 (Student's t-test). (F) SH-SY5Y cells were pre-treated with 5 or 10 μmol/L Ac220 for 2 h and then treated with DMSO, Ac220 (5 or 10 μmol/L), magnolol 100 μmol/L and combined agents for 24 h. Cells were collected, and cell death was quantified using PI live exclusion staining. Data are presented as mean ± SD (n = 3). ∗∗∗P < 0.001 (one-way ANOVA). (G) Representative photos of SH-SY5Y xenografted tumors. Mice were intraperitoneally treated with saline (Ctrl), magnolol (35 mg/kg), HCQ (50 mg/kg), Ac220 (1.5 mg/kg), magnolol + HCQ, magnolol + Ac220 for 23 days. Scale bar: 5 mm. (H) Tumor volumes were measured every three or four days following the treatment in (F). Values are represented as mean ± SD from 6 tumors in each group. ∗∗P < 0.01, ∗∗∗P < 0.001 (two-way ANOVA). (I) Tumor weights were analyzed after 23 days' treatment in (F). Values are represented as mean ± SD from 6 tumors in each group. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (Student's t-test).