Azithromycin enhances anticancer activity of TRAIL by inhibiting autophagy and up‐regulating the protein levels of DR4/5 in colon cancer cells in vitro and in vivo

• Published in: Cancer communications (London, England) Vol. 38; no. 1; pp. 1 - 13
• Main Authors: Qiao, Xinran, Wang, Xiaofei, Shang, Yue, Li, Yi, Chen, Shu‐zhen
• Format: Journal Article
• Language: English
• Published: London BioMed Central 03.07.2018; Wiley
• Subjects: Animals; Anti-Bacterial Agents - pharmacology; Apoptosis; Autophagy; Autophagy - drug effects; Autophagy - genetics; Azithromycin; Azithromycin - pharmacology; Cell Line, Tumor; Colon cancer; Colonic Neoplasms - drug therapy; Colonic Neoplasms - metabolism; Colonic Neoplasms - pathology; Drug Synergism; HCT116 Cells; Humans; Mice, Inbred BALB C; Mice, Nude; Microtubule-Associated Proteins - genetics; Microtubule-Associated Proteins - metabolism; Original; Receptors, TNF-Related Apoptosis-Inducing Ligand - genetics; Receptors, TNF-Related Apoptosis-Inducing Ligand - metabolism; RNA Interference; TNF-Related Apoptosis-Inducing Ligand - pharmacology; TRAIL; Up-Regulation - drug effects; Xenograft Model Antitumor Assays; Colon cancer; TRAIL; Autophagy; Azithromycin; Apoptosis
• ISSN: 2523-3548; 2523-3548
• DOI: 10.1186/s40880-018-0309-9

Background Azithromycin is a member of macrolide antibiotics, and has been reported to inhibit the proliferation of cancer cells. However, the underlying mechanisms are not been fully elucidated. Tumor necrosis factor‐related apoptosis‐inducing ligand (TRAIL) selectively targets tumor cells without damaging healthy cells. In the present study, we examined whether azithromycin is synergistic with TRAIL, and if so, the underlying mechanisms in colon cancers. Methods HCT‐116, SW480, SW620 and DiFi cells were treated with azithromycin, purified TRAIL, or their combination. A sulforhoddamine B assay was used to examine cell survival. Apoptosis was examined using annexin V‐FITC/PI staining, and autophagy was observed by acridine orange staining. Western blot analysis was used to detect protein expression levels. In mechanistic experiments, siRNAs were used to knockdown death receptors (DR4, DR5) and LC‐3B. The anticancer effect of azithromycin and TRAIL was also examined in BALB/c nude mice carrying HCT‐116 xenografts. Results Azithromycin decreased the proliferation of HCT‐116 and SW480 cells in a dose‐dependent manner. Combination of azithromycin and TRAIL inhibited tumor growth in a manner that could not be explained by additive effects. Azithromycin increased the expressions of DR4, DR5, p62 and LC‐3B proteins and potentiated induction of apoptosis by TRAIL. Knockdown of DR4 and DR5 with siRNAs increased cell survival rate and decreased the expression of cleaved‐PARP induced by the combination of azithromycin and TRAIL. LC‐3B siRNA and CQ potentiated the anti‐proliferation activity of TRAIL alone, and increased the expressions of DR4 and DR5. Conclusion The synergistic antitumor effect of azithromycin and TRAIL mainly relies on the up‐regulations of DR4 and DR5, which in turn result from LC‐3B‐involved autophagy inhibition.