Fig. 5: Anticancer effects of penfluridol via inducing autophagosome accumulation in an A549 orthotopic graft model.

Luciferase-tagged A549 cells were orthotopically implanted into the left lateral thorax of NOD/SCID mice. After tumor cell injection for 1 week, mice were orally administered penfluridol (5 or 10 mg/kg) or vehicle 5 days/week for 5 consecutive weeks. a Left panel: xenogen IVIS spectrum bioluminescence imaging of orthotopic lung tumor growth. Right panel: quantitative analysis of Xenogen imaging signal intensity (photons/s/cm²/steradian) at the indicated time points. *p < 0.05; ***p < 0.001 compared with the vehicle control group. b Left panel: cancer metastasis from the left lung to the right lung was imaged by bioluminescence. Right panel: signal intensities from primary tumors (left lung) and metastatic tumors (right lung) were bioluminescently captured at the end of the study, with the mean signal for each group indicated. *p < 0.05, **p < 0.01, ***p < 0.001 compared with the control group. ^##p < 0.01 compared with the 5 mg/kg penfluridol-treated group. Upper panel: cancer distal metastasis, including liver (c) and pancreas (d) metastasis, was imaged with bioluminescence at the end of the study. Lower panel: liver (c) and pancreas (d) metastatic signal intensities were bioluminescently captured with the mean signal for each group indicated. *p < 0.05, **p < 0.01, ***p < 0.001 compared with the control group. A549 orthotopic tumors treated with vehicle or penfluridol were isolated to detect light chain 3 (LC3) conversion by a western blot analysis (e) and autophagosome formation by IHC staining of LC3 (f). Quantitative results of LC3B-II proteins were normalized to α-tubulin levels. LC3 dot-like staining is indicated by an arrow. (Left: original magnification ×400; Right: original magnification ×1000). g Kaplan–Meier analysis of LC3B gene expressions in lung cancer tissues (GSE30219)