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Primary research
Ming Zhang
Department of Otorhinolaryngology, Eye & ENT Hospital, Fudan University, Shanghai, China, 200031
Corresponding Author
ORCiD: https://orcid.org/0000-0003-2909-8446
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Objective: We explored the antitumor effect of dual PI3K/mTOR inhibitor combined with autophagy suppression on laryngeal squamous cell carcinoma (LSCC) and its underlying mechanism.
Methods: Hep-2 and AMC-HN-8 cell lines were treated with the Akt inhibitor LY294002, mTOR inhibitor rapamycin, and dual inhibitor NVP-BEZ235 separately. The biological characteristics of in vitro proliferation, cell cycle, apoptosis, migration, invasion, and autophagy were analyzed, and the expression levels of PI3K/Akt/mTOR pathway-related proteins were also measured. The in vivo effects of NVP-BEZ235 combined with inhibition of autophagy using pharmacological inhibitor was further assessed.
Results: Compared with Akt or mTOR inhibitor, NVP-BEZ235 had the most significant biological effects on LSCC cells. When combined with various autophagy inhibitors, along with siRNA against ATG7, NVP-BEZ235 showed a synergic antitumor effect in LSCC through increasing cell apoptosis and death both in vitro and vivo.
Conclusion: NVP-BEZ235 exerted stronger antitumor effects on LSCC, especially when combined with the autophagy inhibitor, providing convincing experimental data for new molecular targeted therapy for LSCC.
Keywords
laryngeal squamous cell carcinoma (LSCC), PI3K/mTOR inhibitors, autophagy
Figure 1
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This is a list of supplementary files associated with this preprint. Click to download.
- Fig.S1.tif
Fig.S1. MTT assay in HEP-2 and AMC-HN-8 cell lines after treatment with different concentrations of LY294002 (1, 5, 10, 20, 40, 80, 160 μM), rapamycin (1, 5, 10, 20, 40, 80, 160 μM), and NVP-BEZ235 (0.1, 0.5, 1, 2, 4, 8, 16 μM) for 72 h.
- Fig.S2.tif
Fig.S2. Fluorescence micrographs of acridine orange-stained HEP-2 and AMC-HN-8 cell lines after culture in the absence (control) and presence of three different inhibitors.
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A new preprint design is coming!
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This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Primary research
Ming Zhang
Department of Otorhinolaryngology, Eye & ENT Hospital, Fudan University, Shanghai, China, 200031
Corresponding Author
ORCiD: https://orcid.org/0000-0003-2909-8446
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 07 Jun, 2021
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Cancer Biology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Objective: We explored the antitumor effect of dual PI3K/mTOR inhibitor combined with autophagy suppression on laryngeal squamous cell carcinoma (LSCC) and its underlying mechanism.
Methods: Hep-2 and AMC-HN-8 cell lines were treated with the Akt inhibitor LY294002, mTOR inhibitor rapamycin, and dual inhibitor NVP-BEZ235 separately. The biological characteristics of in vitro proliferation, cell cycle, apoptosis, migration, invasion, and autophagy were analyzed, and the expression levels of PI3K/Akt/mTOR pathway-related proteins were also measured. The in vivo effects of NVP-BEZ235 combined with inhibition of autophagy using pharmacological inhibitor was further assessed.
Results: Compared with Akt or mTOR inhibitor, NVP-BEZ235 had the most significant biological effects on LSCC cells. When combined with various autophagy inhibitors, along with siRNA against ATG7, NVP-BEZ235 showed a synergic antitumor effect in LSCC through increasing cell apoptosis and death both in vitro and vivo.
Conclusion: NVP-BEZ235 exerted stronger antitumor effects on LSCC, especially when combined with the autophagy inhibitor, providing convincing experimental data for new molecular targeted therapy for LSCC.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
This is a list of supplementary files associated with this preprint. Click to download.
- Fig.S1.tif
Fig.S1. MTT assay in HEP-2 and AMC-HN-8 cell lines after treatment with different concentrations of LY294002 (1, 5, 10, 20, 40, 80, 160 μM), rapamycin (1, 5, 10, 20, 40, 80, 160 μM), and NVP-BEZ235 (0.1, 0.5, 1, 2, 4, 8, 16 μM) for 72 h.
- Fig.S2.tif
Fig.S2. Fluorescence micrographs of acridine orange-stained HEP-2 and AMC-HN-8 cell lines after culture in the absence (control) and presence of three different inhibitors.
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