2,3 ',4,4 ',5-Pentachlorobiphenyl Induced Thyrocyte Autophagy by Promoting Calcium Influx via Store-Operated Ca2+ Entry
2,3 ',4,4 ',5-Pentachlorobiphenyl Induced Thyrocyte Autophagy by Promoting Calcium Influx via Store-Operated Ca2+ Entry
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DOI：
10.1093/toxsci/kfaa116
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Duan Y
中科院分区：
文献类型：
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作者：
Wang L;Xu W;Zhou Q;Xu B;Sheng Y;Sun M;Chen H;Wang Y;Ding G;Duan Y
PCB 118是一种2，3 ′，4，4 ′，5-五氯联苯，可破坏甲状腺超微结构，诱导甲状腺细胞自噬。以前，我们报道了PCB 118在体内和体外促进自噬体形成，但更多的细节仍有待揭示。为了探索PCB 118调节甲状腺细胞自噬的潜在机制，Fischer大鼠甲状腺细胞系-5（FRTL-5）细胞暴露于不同剂量的PCB 118（0、0.25、2.5和25 nM）0-48 h。自噬相关蛋白P62、BECLIN 1和LC 3的Western印迹分析表明，PCB 118以剂量和时间依赖性方式诱导自噬形成。激光扫描共聚焦显微镜和流式细胞术显示PCB 118处理导致细胞内钙浓度（[Ca 2 +]i）呈时间和剂量依赖性增加。此外，PCB 118促进了钙库操纵的钙内流（SOCE）通道，随后显著增加了ORAI 1和STIM 1蛋白水平。另一方面，在FRTL-5细胞中，PCB 118通过III类β微管蛋白（TUBB 3）/死亡相关蛋白激酶2（DAPK 2）/肌球蛋白调节轻链（MRLC）/自噬相关9A（ATG 9A）途径诱导甲状腺自噬。用SOCE抑制剂SKF 96365预处理降低了胞浆Ca 2+、ORAI 1、STIM 1和BECLIN 1水平以及LC 3 II/LC 3 I比率，同时增加了P62表达。SKF 96365对PCB 118处理的FRTL-5细胞中TUBB 3/DAPK 2/MRLC/ATG 9A通路也有抑制作用。我们的研究结果提供了证据表明，PCB 118可能通过TUBB 3相关的信号通路诱导甲状腺自噬，这些作用可能是通过SOCE通道的钙内流调节。
PCB118, a 2,3′,4,4′,5-pentachlorobiphenyl, has been shown to destroy thyroidal ultrastructure and induce thyrocyte autophagy. Previously, we reported that PCB118 promoted autophagosome formationin vivoandin vitro, but more details remain to be revealed. To explore the underlying mechanism by which PCB118 regulates thyrocyte autophagy, Fischer rat thyroid cell line-5 (FRTL-5) cells were exposed to different doses of PCB118 at 0, 0.25, 2.5, and 25 nM for 0–48 h. Western blot analysis of autophagy-related proteins P62, BECLIN1, and LC3 demonstrated that PCB118 induced autophagy formation in dose- and time-dependent manner. Moreover, laser scanning confocal microscopy and flow cytometry showed PCB118 treatment led to time- and dose-dependent increase in intracellular calcium concentration ([Ca2+]i). Additionally, PCB118 promoted store-operated Ca2+entry (SOCE) channel followed by significant increase of ORAI1 and STIM1 protein levels. On the other hand, PCB118 induced thyroidal autophagy via class III β‐tubulin (TUBB3)/death-associated protein kinase 2 (DAPK2)/myosin regulatory light chain (MRLC)/autophagy-related 9A (ATG9A) pathway in FRTL-5 cells. Pretreatment with SOCE inhibitor SKF96365 reduced cytosolic Ca2+, ORAI1, STIM1, and BECLIN1 levels as well as LC3 II/LC3 I ratio, while increased P62 expression. SKF96365 also inhibited TUBB3/DAPK2/MRLC/ATG9A pathway in FRTL-5 cells treated by PCB118. Our results provide evidence that PCB118 may induce thyroidal autophagy through TUBB3-related signaling pathway, and these effects are likely to be regulated by calcium influx via SOCE channel.