CWHM-1008 Induces Apoptosis and Protective Autophagy through the Akt/mTOR Axis in LUAD Cells

Figure 3

CWHM-1008 stimulates autophagy by blocking the Akt/mTOR axis in lung adenocarcinoma cells. (a) H358 (left) and A549 (right) cells were transiently infected with GFP-RFP-LC3 adenovirus. In addition, H358 and A549 cells were treated with various concentrations of CWHM-1008 (0, 20, 30, or 40 µM) for 24 h. Scale bars, 20 µm. (b) Total number of LC3-GFP and LC3-RFP puncta per cell. (c, d) Levels of autophagy-related protein LC3, ATG7, beclin-1, and P62 in H358 and A549 cells treated with various concentrations of CWHM-1008 for 24 h were analyzed by western blotting. (e, f) Levels of RAC-alpha serine/threonine-protein kinase (Akt), phospho-Akt, mammalian target of rapamycin (mTOR), and phosphorylated-mTOR in H358 and A549 cells treated with the indicated concentrations of CWHM-1008 for 24 h were analyzed by western blotting. Data are presented as the mean ± SD, n = 3. , , and ; ns, not significant.

(a)

(b)

(c)

(d)

(e)

(f)