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Synergistic Effect and Molecular Mechanism of Homoharringtonine and Bortezomib on SKM-1 Cell Apoptosis.

Zhang J, Chen B, Wu T, Wang Q, Zhuang L, Zhu C, Fan N, Qing W, Ma Y, Xu X - PLoS ONE (2015)

Bottom Line: Simultaneous exposure to HHT and Bortezomib (10.4:1) resulted in a significant reduction of cell proliferation in SKM-1 cells (P < 0.05).HHT and Bortezomib synergistically inhibit SKM-1 cell proliferation and induce apoptosis in vitro.Inhibition of Akt and NF-κB pathway signaling contribute to molecular mechanism of HHT and Bortezomib. miR-3151 abundance is implicated in SKM-1 cell viability, cell proliferation and p53 protein expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai, China.

ABSTRACT

Background: Myelodysplastic syndromes (MDS) are clonal marrow stem-cell disorders with a high risk of progression to acute myeloid leukemia (AML). Treatment options are limited and targeted therapies are not available for MDS. In the present study, we investigated the cytotoxicity and the molecular mechanism of Homoharringtonine (HHT) and Bortezomib towards high-risk MDS cell line SKM-1 in vitro and the role of miR-3151 was first evaluated in SKM-1 cells.

Methods: SKM-1 cells were treated with different concentrations of HHT or Bortezomib, and cell viability was analyzed with CCK-8 assay. The influence on cell proliferation, cell cycle distribution and the percentage of apoptosis cells were analyzed by flow cytometry. Calcusyn software was used to calculate combination index (CI) values. Western blot was used to analysis phosphorylation of Akt and nuclear NF-κB protein expression in SKM-1 cells. Mature miR-3151 level and p53 protein level were detected after HHT or Bortezomib treatment. The cell proliferation and p53 protein level were reassessed in SKM-1 cells infected with lentivirus to overexpress miR-3151.

Results: Simultaneous exposure to HHT and Bortezomib (10.4:1) resulted in a significant reduction of cell proliferation in SKM-1 cells (P < 0.05). Cell cycle arrest at G0/G1 and G2/M phase was observed (P < 0.05). HHT and Bortezomib synergistically induced cell apoptosis by regulating members of caspase 9, caspase 3 and Bcl-2 family (P < 0.01). The mechanisms of the synergy involved Akt and NF-κB signaling pathway inhibition, downregulation of mature miR-3151 and increment of downstream p53 protein level. Overexpression of miR-3151 promoted cell proliferation and inhibited p53 protein expression in SKM-1 (P < 0.01).

Conclusions: HHT and Bortezomib synergistically inhibit SKM-1 cell proliferation and induce apoptosis in vitro. Inhibition of Akt and NF-κB pathway signaling contribute to molecular mechanism of HHT and Bortezomib. miR-3151 abundance is implicated in SKM-1 cell viability, cell proliferation and p53 protein expression.

No MeSH data available.


Related in: MedlinePlus

Effects of HHT and Bortezomib on caspases and Bcl-2 family proteins.HHT and Bortezomib simultaneously actived caspase 9 and caspase 3, induced the up-regulation of Bax protein and the down-regulation of Bcl-2 protein after 72 h exposure. *P < 0.05, #P < 0.01 vs untreated control.
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pone.0142422.g005: Effects of HHT and Bortezomib on caspases and Bcl-2 family proteins.HHT and Bortezomib simultaneously actived caspase 9 and caspase 3, induced the up-regulation of Bax protein and the down-regulation of Bcl-2 protein after 72 h exposure. *P < 0.05, #P < 0.01 vs untreated control.

Mentions: Western blotting was used to evaluate the expression of caspase 9, caspase 3, Bcl-2 and Bax in SKM-1 cells after treatment with HHT, Bortezomib or combination of HHT and Bortezomib for 72 h. Caspase 9 and caspase 3 are important members of the caspase family. Cleaved caspase 9 further processes caspase 3 to initiate a caspase cascade leading to apoptosis. The results showed that 11.55 nM HHT or 1.12 nM Bortezomib significantly increased cleavage activation of caspase 9 and caspase 3, inhibited anti-apoptotic protein Bcl-2 expression and upregulated pro-apoptotic protein Bax. The effect of drugs was more obvious when HHT: Bortezomib was 10.4:1 (Fig 5).


Synergistic Effect and Molecular Mechanism of Homoharringtonine and Bortezomib on SKM-1 Cell Apoptosis.

Zhang J, Chen B, Wu T, Wang Q, Zhuang L, Zhu C, Fan N, Qing W, Ma Y, Xu X - PLoS ONE (2015)

Effects of HHT and Bortezomib on caspases and Bcl-2 family proteins.HHT and Bortezomib simultaneously actived caspase 9 and caspase 3, induced the up-regulation of Bax protein and the down-regulation of Bcl-2 protein after 72 h exposure. *P < 0.05, #P < 0.01 vs untreated control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4636319&req=5

pone.0142422.g005: Effects of HHT and Bortezomib on caspases and Bcl-2 family proteins.HHT and Bortezomib simultaneously actived caspase 9 and caspase 3, induced the up-regulation of Bax protein and the down-regulation of Bcl-2 protein after 72 h exposure. *P < 0.05, #P < 0.01 vs untreated control.
Mentions: Western blotting was used to evaluate the expression of caspase 9, caspase 3, Bcl-2 and Bax in SKM-1 cells after treatment with HHT, Bortezomib or combination of HHT and Bortezomib for 72 h. Caspase 9 and caspase 3 are important members of the caspase family. Cleaved caspase 9 further processes caspase 3 to initiate a caspase cascade leading to apoptosis. The results showed that 11.55 nM HHT or 1.12 nM Bortezomib significantly increased cleavage activation of caspase 9 and caspase 3, inhibited anti-apoptotic protein Bcl-2 expression and upregulated pro-apoptotic protein Bax. The effect of drugs was more obvious when HHT: Bortezomib was 10.4:1 (Fig 5).

Bottom Line: Simultaneous exposure to HHT and Bortezomib (10.4:1) resulted in a significant reduction of cell proliferation in SKM-1 cells (P < 0.05).HHT and Bortezomib synergistically inhibit SKM-1 cell proliferation and induce apoptosis in vitro.Inhibition of Akt and NF-κB pathway signaling contribute to molecular mechanism of HHT and Bortezomib. miR-3151 abundance is implicated in SKM-1 cell viability, cell proliferation and p53 protein expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai, China.

ABSTRACT

Background: Myelodysplastic syndromes (MDS) are clonal marrow stem-cell disorders with a high risk of progression to acute myeloid leukemia (AML). Treatment options are limited and targeted therapies are not available for MDS. In the present study, we investigated the cytotoxicity and the molecular mechanism of Homoharringtonine (HHT) and Bortezomib towards high-risk MDS cell line SKM-1 in vitro and the role of miR-3151 was first evaluated in SKM-1 cells.

Methods: SKM-1 cells were treated with different concentrations of HHT or Bortezomib, and cell viability was analyzed with CCK-8 assay. The influence on cell proliferation, cell cycle distribution and the percentage of apoptosis cells were analyzed by flow cytometry. Calcusyn software was used to calculate combination index (CI) values. Western blot was used to analysis phosphorylation of Akt and nuclear NF-κB protein expression in SKM-1 cells. Mature miR-3151 level and p53 protein level were detected after HHT or Bortezomib treatment. The cell proliferation and p53 protein level were reassessed in SKM-1 cells infected with lentivirus to overexpress miR-3151.

Results: Simultaneous exposure to HHT and Bortezomib (10.4:1) resulted in a significant reduction of cell proliferation in SKM-1 cells (P < 0.05). Cell cycle arrest at G0/G1 and G2/M phase was observed (P < 0.05). HHT and Bortezomib synergistically induced cell apoptosis by regulating members of caspase 9, caspase 3 and Bcl-2 family (P < 0.01). The mechanisms of the synergy involved Akt and NF-κB signaling pathway inhibition, downregulation of mature miR-3151 and increment of downstream p53 protein level. Overexpression of miR-3151 promoted cell proliferation and inhibited p53 protein expression in SKM-1 (P < 0.01).

Conclusions: HHT and Bortezomib synergistically inhibit SKM-1 cell proliferation and induce apoptosis in vitro. Inhibition of Akt and NF-κB pathway signaling contribute to molecular mechanism of HHT and Bortezomib. miR-3151 abundance is implicated in SKM-1 cell viability, cell proliferation and p53 protein expression.

No MeSH data available.


Related in: MedlinePlus