Pang XL, He G, Liu YB, Wang Y, Zhang B. group, BEZ235 (inhibitors of PI3K/AKT/mTOR pathway) group and tunicamycin + BEZ235 group. Cell apoptosis was recognized by circulation cytometry. Autophagy was observed by fluorescence microscopy and circulation cytometry. Western blotting was used to detect the expressions of ERS-related proteins, autophagy-related proteins, apoptosis-related proteins and PI3K/AKT/mTOR pathway-related proteins. Conclusions Our findings provide evidence the activation of ERS could promote autophagy and apoptosis and reverse chemoresistance of human being SCLC cells by inhibiting the PI3K/AKT/mTOR pathway. < 0.05) (Figure ?(Figure1A).1A). Consequently, NCI-H446 and H69 cells were selected for further experiments. Open in a separate window Number 1 Effects of different doses of tunicamycin within the viability of human being small cell lung malignancy (SCLC) cells(A) assessment of the viability of five SCLC cell lines treated with 5 ug/mL tunicamycin for 24 h; *stands for < 0.05 in comparison with NCI-H446 cells; #stands for < 0.05 in comparison with H69 cells; (B) changes in NCI-H466 cell viability after treated with different doses of tunicamycin; (C), changes in H69 cell viability after treated with different doses of tunicamycin; *stands for < 0.05 in comparison with 0 ug/mL tunicamycin. The effects of tunicamycin within the viability of NCI-H446 and H69 cells were inside a dose-dependent and time-dependent manner. With the increasing doses of tunicamycin, the effects of the tunicamycin within the viability of NCI-H446 and H69 cells were increased continually (all < 0.05) (Figure ?(Number1B1B and Apogossypolone (ApoG2) ?and1C).1C). The IC50 ideals after 24 h of tunicamycin treatment on NCI-H446 and H69 cells were 3.01 0.14 g/mL and 2.94 0.16 g/mL, respectively. Effects of different doses of tunicamycin within the expressions of ESR-related proteins Apogossypolone (ApoG2) and PI3K/AKT/mTOR signaling pathway-related proteins in NCI-H446 and H69 cells Tunicamycin can activate ERS and up-regulate the expressions TSHR of ERS-related Apogossypolone (ApoG2) proteins (PERK, eIF2 and CHOP) inside a dose-dependent and time-dependent manner (all < 0.05) (Figure ?(Figure2).2). The tunicamycin can inhibit PI3K/AKT/mTOR signaling pathway and down-regulate the expressions of p-PI3K, p-AKT and p-mTOR, and the effects were increased significantly with the increasing doses of tunicamycin (all < 0.05). However, the expressions of PI3K, AKT, or mTOR showed no changes in NCI-H446 and H69 cells at 24 after tunicamycin treatment (Number ?(Figure3).3). These results suggest that the activation of ERS could inhibit the PI3K/AKT/mTOR signaling pathway. Open in a separate window Number 2 Effects of different doses of tunicamycin within the activation of endoplasmic reticulum stress (ERS) in NCI-H446 and H69 cells(A), ERS-related protein expressions in NCI-H446 cells after treated with different doses of tunicamycin for 24 h; (B), ERS-related protein expressions in H69 cells after treated with different doses of tunicamycin for 24 h; *stands for < 0.05 in comparison with 0 ug/mL tunicamycin. Open in a separate window Number 3 Effects of different doses of tunicamycin within the PI3K/AKT/mTOR signaling pathway in NCI-H446 and H69 cells(A), the expressions of PI3K/AKT/mTOR signaling pathway-related proteins in NCI-H446 cells after treated with different doses of tunicamycin for 24 h; (B), the expressions of PI3K/AKT/mTOR-related proteins in H69 cells after treated with different doses of tunicamycin for 24 h; *stands for < 0.05 in comparison with 0 ug/mL tunicamycin. Effects of different doses of tunicamycin within the autophagy and apoptosis of NCI-H446 and H69 cells Tunicamycin can induce autophagy of NCI-H446 and H69 cells and regulate the expressions of autophagy-related proteins. With the increasing doses of tunicamycin, the protein expressions of LC3, LC3-II and Beclin1 improved continuously, but the protein expressions of.