Xin Pengliang, Xu Wenqian, Zhu Xiongpeng, Li Chuntuan, Zheng Yan, Zheng Tingjin, Cheng Wenzhao, Peng Qunyi
Department of Haematology, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, People's Republic of China.
Central Laboratory, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, People's Republic of China.
Cancer Manag Res. 2019 Aug 22;11:7933-7951. doi: 10.2147/CMAR.S204472. eCollection 2019.
To investigate the effects of BEZ235 on chronic myeloid leukemia (CML) cells.
MTS assay was used to detect the proliferation of CML cells. The proteins expression were detected by Western blot assay. The effects of BEZ235 on autophagy in CML cells were verified through transmission electron microscopy and evaluated by laser confocal microscopy. Annexin V-FITC/PI double staining flow cytometry was used to detect apoptosis. A xenograft model was established to observe the therapeutic effect of BEZ235 in vivo.
BEZ235 could inhibit the proliferation of CML cells; CQ and 3-MA could increase the proliferation inhibition and Z-VAD-FMK can reduce the proliferation inhibition of BEZ235 on CML cells (<0.05). Results of TEM showed that the autophagosomes of CML cells treated with BEZ235 increased (<0.05). The results by confocal microscopy showed that the autophagic activity of K562 cells increased with BEZ235 treatment. When BEZ235 combined with CQ, BEZ235-induced autophagic flow was blocked. FCM results showed that BEZ235 could induces apoptosis in CML cells. Z-VAD-FMK could decrease the apoptosis of CML cells induced by BEZ235. CQ increased the apoptosis of CML cells induced by BEZ235 (<0.05). Western blot showed that BEZ235 inhibited the phosphorylation of AKT and S6K. BEZ235 alone could upregulate the expression of cleaved caspase-3 and LC3II. When combined with Z-VAD-FMK, the expression of cleaved caspase-3 was lower than that of BEZ235 alone. When combined with CQ, the expression of cleaved caspase-3 and LC3II were higher than those of BEZ235 alone (<0.05). BEZ235 could inhibit the growth of xenografts of CML cell line.
BEZ235 can inhibit the proliferation of CML cells, induce apoptosis, and enhance autophagy activity. It induces protective autophagy. The combination of CQ can enhance the apoptosis and proliferation inhibition of CML cells induced by BEZ235.
研究BEZ235对慢性髓性白血病（CML）细胞的作用。
采用MTS法检测CML细胞的增殖。通过蛋白质免疫印迹法检测蛋白表达。通过透射电子显微镜验证BEZ235对CML细胞自噬的影响，并通过激光共聚焦显微镜进行评估。采用膜联蛋白V-异硫氰酸荧光素/碘化丙啶双染流式细胞术检测细胞凋亡。建立异种移植模型以观察BEZ235在体内的治疗效果。
BEZ235可抑制CML细胞的增殖；氯喹（CQ）和3-甲基腺嘌呤（3-MA）可增强增殖抑制作用，而Z-VAD-FMK可降低BEZ235对CML细胞的增殖抑制作用（P<0.05）。透射电镜结果显示，经BEZ235处理的CML细胞自噬体增多（P<0.05）。共聚焦显微镜结果显示，K562细胞经BEZ235处理后自噬活性增强。当BEZ235与CQ联合使用时，BEZ235诱导的自噬流被阻断。流式细胞术结果显示，BEZ235可诱导CML细胞凋亡。Z-VAD-FMK可降低BEZ235诱导的CML细胞凋亡。CQ可增强BEZ235诱导的CML细胞凋亡（P<0.05）。蛋白质免疫印迹显示，BEZ235抑制AKT和S6K的磷酸化。单独使用BEZ235可上调裂解的半胱天冬酶-3（cleaved caspase-3）和微管相关蛋白轻链3-II（LC3II）的表达。与Z-VAD-FMK联合使用时，裂解的半胱天冬酶-3的表达低于单独使用BEZ235时。与CQ联合使用时，裂解的半胱天冬酶-3和LC3II的表达高于单独使用BEZ235时（P<0.05）。BEZ235可抑制CML细胞系异种移植瘤的生长。
BEZ235可抑制CML细胞的增殖，诱导细胞凋亡，并增强自噬活性。它诱导保护性自噬。CQ联合使用可增强BEZ235诱导的CML细胞凋亡和增殖抑制作用。
