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Rapamycin Enhances the Anti-Cancer Effect of Dasatinib by Suppressing Src/PI3K/mTOR Pathway in NSCLC Cells.

Chen B, Xu X, Luo J, Wang H, Zhou S - PLoS ONE (2015)

Bottom Line: In this study, we found that Rapamycin dramatically enhanced Dasatinib-induced cell growth inhibition and cell cycle G1 arrest in human lung adenocarcinoma A549 cells without affecting apoptosis.The synergistic effects were consistently correlated with the up-regulation of cyclin-dependent kinases inhibitor proteins, including p16, p19, p21, and p27, as well as the repression of Cdk4 expression and nuclear translocation.Restraining Src and mTOR with small interfering RNA in A549 cells further confirmed that the Src/PI3K/mTOR Pathway played a crucial role in enhancing the anticancer effect of Dasatinib.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, Shanghai Pulmonary Hospital, Shanghai, China; School of Medicine Cancer Institute, Tongji University, Shanghai, China.

ABSTRACT
Src and the mammalian target of rapamycin (mTOR) signaling are commonly activated in non-small cell lung cancer (NSCLC) and hence potential targets for chemotherapy. Although the combined use of Src inhibitor Dasatinib with other chemotherapeutic agents has shown superior efficacy for cancer treatment, the mechanisms that lead to enhanced sensitivity of Dasatinib are not completely understood. In this study, we found that Rapamycin dramatically enhanced Dasatinib-induced cell growth inhibition and cell cycle G1 arrest in human lung adenocarcinoma A549 cells without affecting apoptosis. The synergistic effects were consistently correlated with the up-regulation of cyclin-dependent kinases inhibitor proteins, including p16, p19, p21, and p27, as well as the repression of Cdk4 expression and nuclear translocation. Mechanistic investigations demonstrated that FoxO1/FoxO3a and p70S6K/4E-BP1, the molecules at downstream of Src-PI3K-Akt and mTOR signaling, were significantly suppressed by the combined use of Dasatinib and Rapamycin. Restraining Src and mTOR with small interfering RNA in A549 cells further confirmed that the Src/PI3K/mTOR Pathway played a crucial role in enhancing the anticancer effect of Dasatinib. In addition, this finding was also validated by a series of assays using another two NSCLC cell lines, NCI-H1706 and NCI-H460. Conclusively, our results suggested that the combinatory application of Src and mTOR inhibitors might be a promising therapeutic strategy for NSCLC treatment.

No MeSH data available.


Related in: MedlinePlus

Rapamycin enhanced the inhibiting effect of Dasatinib on invasion and migration in A549 cells.(A) Representative images (upper panel) and quantification (lower panel) of invading cells. As detailed by “cell invasion assay” in the “Methods”, A549 cells were treated with DMSO (0.1%), Dasatinib (10 nM) alone or in combination with Rapamycin (100 nM) for 24 h. The numbers of invading cells were counted under microscope and the data was presented as mean ± SD from three independent experiments, * p < 0.05, ** p < 0.01. (B) Representative images of wound healing assays in A549 cells that were treated with DMSO (0.1%), Dasatinib (10 nM) or Rapamycin (100 nM) for 18 h. (C) The expression of MMP-2/9 and E-cadherin determined by western blotting in A549 cells that were treated with DMSO (0.1%), Dasatinib (10 nM) or Rapamycin (100 nM) for 24 h.
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pone.0129663.g005: Rapamycin enhanced the inhibiting effect of Dasatinib on invasion and migration in A549 cells.(A) Representative images (upper panel) and quantification (lower panel) of invading cells. As detailed by “cell invasion assay” in the “Methods”, A549 cells were treated with DMSO (0.1%), Dasatinib (10 nM) alone or in combination with Rapamycin (100 nM) for 24 h. The numbers of invading cells were counted under microscope and the data was presented as mean ± SD from three independent experiments, * p < 0.05, ** p < 0.01. (B) Representative images of wound healing assays in A549 cells that were treated with DMSO (0.1%), Dasatinib (10 nM) or Rapamycin (100 nM) for 18 h. (C) The expression of MMP-2/9 and E-cadherin determined by western blotting in A549 cells that were treated with DMSO (0.1%), Dasatinib (10 nM) or Rapamycin (100 nM) for 24 h.

Mentions: To further confirm the enhancing effect of Rapamycin on the anticancer activity of Dasatinib, we examined the invasion and migration of A549 cells under various treatments. As shown in Fig 5A, the invasive ability through Matrigel-coated filters of cells treated by Dasatinib was significantly decreased in comparison with the control cells. The co-treatment of Dasatinib with Rapamycin further decreased the invasion percent of A549 cells by nearly 50%, whereas Rapamycin alone did not exhibit clear suppression on cell invasion when compared to the control.


Rapamycin Enhances the Anti-Cancer Effect of Dasatinib by Suppressing Src/PI3K/mTOR Pathway in NSCLC Cells.

Chen B, Xu X, Luo J, Wang H, Zhou S - PLoS ONE (2015)

Rapamycin enhanced the inhibiting effect of Dasatinib on invasion and migration in A549 cells.(A) Representative images (upper panel) and quantification (lower panel) of invading cells. As detailed by “cell invasion assay” in the “Methods”, A549 cells were treated with DMSO (0.1%), Dasatinib (10 nM) alone or in combination with Rapamycin (100 nM) for 24 h. The numbers of invading cells were counted under microscope and the data was presented as mean ± SD from three independent experiments, * p < 0.05, ** p < 0.01. (B) Representative images of wound healing assays in A549 cells that were treated with DMSO (0.1%), Dasatinib (10 nM) or Rapamycin (100 nM) for 18 h. (C) The expression of MMP-2/9 and E-cadherin determined by western blotting in A549 cells that were treated with DMSO (0.1%), Dasatinib (10 nM) or Rapamycin (100 nM) for 24 h.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4465694&req=5

pone.0129663.g005: Rapamycin enhanced the inhibiting effect of Dasatinib on invasion and migration in A549 cells.(A) Representative images (upper panel) and quantification (lower panel) of invading cells. As detailed by “cell invasion assay” in the “Methods”, A549 cells were treated with DMSO (0.1%), Dasatinib (10 nM) alone or in combination with Rapamycin (100 nM) for 24 h. The numbers of invading cells were counted under microscope and the data was presented as mean ± SD from three independent experiments, * p < 0.05, ** p < 0.01. (B) Representative images of wound healing assays in A549 cells that were treated with DMSO (0.1%), Dasatinib (10 nM) or Rapamycin (100 nM) for 18 h. (C) The expression of MMP-2/9 and E-cadherin determined by western blotting in A549 cells that were treated with DMSO (0.1%), Dasatinib (10 nM) or Rapamycin (100 nM) for 24 h.
Mentions: To further confirm the enhancing effect of Rapamycin on the anticancer activity of Dasatinib, we examined the invasion and migration of A549 cells under various treatments. As shown in Fig 5A, the invasive ability through Matrigel-coated filters of cells treated by Dasatinib was significantly decreased in comparison with the control cells. The co-treatment of Dasatinib with Rapamycin further decreased the invasion percent of A549 cells by nearly 50%, whereas Rapamycin alone did not exhibit clear suppression on cell invasion when compared to the control.

Bottom Line: In this study, we found that Rapamycin dramatically enhanced Dasatinib-induced cell growth inhibition and cell cycle G1 arrest in human lung adenocarcinoma A549 cells without affecting apoptosis.The synergistic effects were consistently correlated with the up-regulation of cyclin-dependent kinases inhibitor proteins, including p16, p19, p21, and p27, as well as the repression of Cdk4 expression and nuclear translocation.Restraining Src and mTOR with small interfering RNA in A549 cells further confirmed that the Src/PI3K/mTOR Pathway played a crucial role in enhancing the anticancer effect of Dasatinib.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, Shanghai Pulmonary Hospital, Shanghai, China; School of Medicine Cancer Institute, Tongji University, Shanghai, China.

ABSTRACT
Src and the mammalian target of rapamycin (mTOR) signaling are commonly activated in non-small cell lung cancer (NSCLC) and hence potential targets for chemotherapy. Although the combined use of Src inhibitor Dasatinib with other chemotherapeutic agents has shown superior efficacy for cancer treatment, the mechanisms that lead to enhanced sensitivity of Dasatinib are not completely understood. In this study, we found that Rapamycin dramatically enhanced Dasatinib-induced cell growth inhibition and cell cycle G1 arrest in human lung adenocarcinoma A549 cells without affecting apoptosis. The synergistic effects were consistently correlated with the up-regulation of cyclin-dependent kinases inhibitor proteins, including p16, p19, p21, and p27, as well as the repression of Cdk4 expression and nuclear translocation. Mechanistic investigations demonstrated that FoxO1/FoxO3a and p70S6K/4E-BP1, the molecules at downstream of Src-PI3K-Akt and mTOR signaling, were significantly suppressed by the combined use of Dasatinib and Rapamycin. Restraining Src and mTOR with small interfering RNA in A549 cells further confirmed that the Src/PI3K/mTOR Pathway played a crucial role in enhancing the anticancer effect of Dasatinib. In addition, this finding was also validated by a series of assays using another two NSCLC cell lines, NCI-H1706 and NCI-H460. Conclusively, our results suggested that the combinatory application of Src and mTOR inhibitors might be a promising therapeutic strategy for NSCLC treatment.

No MeSH data available.


Related in: MedlinePlus