2,3′,4,4′,5-Pentachlorobiphenyl induced autophagy of the thyrocytes via DAPK2/PKD/VPS34 pathway

2,3 ',4,4 ',5-Pentachlorobiphenyl (PCB118) has been shown to cause thyroidal ultrastructure lesions, but the underlying mechanism remains elusive. This study aimed to elucidate the mechanism by which PCB118 induces the abnormalities of the thyrocytes. Wistar rats were injected intraperitoneally with PCB118 (0, 10, 100 and 1000 mu g/kg/d) for 13weeks, and FRTL-5 cells were treated with PCB118 (0, 0.25, 2.5 and 25nM). Transmission electron microscopy showed typical autophagosomes in the thyroid of PCB118-treated rats. Immunofluorescence staining showed dose-dependent increase of autophagy in FRTL-5 cells exposed to PCB118. In vivo and vitro studies found that Tubulin beta 3 class III (Tubb3) mRNA and protein levels decreased significantly, while Death-associated protein kinase 2 (DAPK2) increased after PCB118 exposure, and the binding between Tubb3 and DAPK2 was enhanced by PCB118 in a dose-dependent manner. Moreover, PCB118 resulted in the upregulation of Protein kinase D (PKD) and downregulation of Phosphatidylinositol 3-kinase (VPS34) in mRNA levels, and the activation of PKD and VPS34 phosphorylation. Additionally, Tubb3 small interfering RNA (siTubb3) suppressed DAPK2 protein expression and PKD phosphorylation in FRTL-5 cells, while VPS34 phosphorylation was inhibited by siPKD. Furthermore, DAPK2, PKD and VPS34 were upregulated by Tubb3 overexpression following PCB118 exposure. Our results demonstrate that low concentrations of PCB118 could promote thyroid autophagy formation and cause the abnormalities in thyroidal ultrastructure, and these effects are likely to be mediated by DAPK2/PKD/VPS34 dependent pathway.