Blue light-induced apoptosis of human promyelocytic leukemia cells via the mitochondrial-mediated signaling pathway

Abstract Objective: To investigate the effect and mechanisms of KPT-330 on cell apoptosis of human leukemia cells. Methods: CCK-8 assay was used to quantify the growth inhibition of cells after exposure to KPT-330 for 24 hours. Fluorescence microscope was used to observe the morphological change of leukemia cell when treated with KPT330 for 24 hours. Annexin V/propidium iodide staining followed by flow cytometric analysis was used to detect apoptosis of leukemia cells. Then Western blot was used to detect the expression level of CRM1 and the activation of apoptosis related protein Caspase-9、Caspase-3 and PARP level. LncRNA microarray was used to analyze the differentially expressed genes from the cluster analyses. Results: The inhibition of cell proliferation of leukemia cells was in a dose-dependent manner and KPT-330 treatment induced expression of CRM1 down-regulated .Abnormal cells were observed under fluorescence microscopy. Cell apoptosis, cell cycle and activation of apoptosis markers demonstrates that KPT-330 induced apoptosis in leukemia cells. LncRNA microarray analysis showed differently expressed mRNAs and lncRNAs in KPT-330 treated HL-60 cells compared with control group. Molecular function analysis showed that KPT-330 induced apoptosis in leukemia cells partially related with Glutathione metabolism and Toll-like receptor signaling pathway. Conclusion: In this study, KPT-330 treatment resulted in inhibition of cell proliferation and induction of apoptosis in leukemia cells. LncRNA microarray analysis showed differentially expressed genes and lncRNAs in KPT-330 treated HL-60 cells and we demonstrated that KPT-330 induced apoptosis in leukemia cells partially related with Glutathione metabolism and Toll-like receptor signaling pathway. These results may provide new clue about molecular mechanism of KPT-330 induced apoptosis; however, underlying details will be required to determine further. Taken together, our findings suggest that for the first time that KPT-330 may act as new candidate's drug for leukemia. Disclosures No relevant conflicts of interest to declare.

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23-Hydroxyursolic Acid Isolated from the Stem Bark of Cussonia bancoensis Induces Apoptosis through Fas/Caspase-8-Dependent Pathway in HL-60 Human Promyelocytic Leukemia Cells

Molecules ◽

10.3390/molecules23123306 ◽

2018 ◽

Vol 23 (12) ◽

pp. 3306 ◽

Cited By ~ 4

Author(s):

Jong-Heon Won ◽

Kyung-Sook Chung ◽

Eun-Young Park ◽

Jeong-Hun Lee ◽

Jung-Hye Choi ◽

...

Keyword(s):

Dna Fragmentation ◽

Molecular Mechanisms ◽

Caspase 3 ◽

Stem Bark ◽

Promyelocytic Leukemia ◽

Leukemia Cells ◽

Caspase 8 ◽

Signaling Complex ◽

Induced Apoptosis ◽

Apoptotic Effects

The natural product 23-hydroxyursolic acid (23-HUA) is a derivative of ursolic acid, which is known to induce cancer cell apoptosis. However, apoptotic effects and mechanisms of 23-HUA have not been well characterized yet. Herein, we investigated the molecular mechanisms of 23-HUA-induced apoptosis in HL-60 human promyelocytic leukemia cells. 23-HUA-treated HL-60 cells showed apoptotic features including internucleosomal DNA condensation and fragmentation as well as externalization of phosphatidylserine residues. 23-HUA induced a series of mitochondrial events including disruption of mitochondrial membrane potential (ΔΨm), cytochrome c and Smac/DIABLO release and loss of balance between pro-apoptotic and anti-apoptotic Bcl-2 proteins in HL-60 cells. In addition, 23-HUA activated caspase-8, caspase-9 and caspase-3. Pretreatment with a broad caspase inhibitor (z-VAD-fmk), a caspase-3 inhibitor (z-DEVD-fmk), and a caspase-8 inhibitor (z-IETD-fmk) significantly attenuated 23-HUA-induced DNA fragmentation. After 23-HUA-induced apoptosis, proteins expression levels of FasL, Fas and FADD constituting the death-inducing signaling complex (DISC) were upregulated in HL-60 cells. Moreover, transfection with Fas or FADD siRNA significantly blocked 23-HUA-induced DNA fragmentation and caspases activation. Taken together, these findings indicate that 23-HUA induces apoptosis in HL-60 human promyelocytic leukemia cells through formation of DISC and caspase-8 activation leading to loss of ΔΨm and caspase-3 activation.

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