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Interference of mTOR expression enhances sensitivity of esophageal squamous cell carcinoma EC9706 cells to rapamycin
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Abstract:
Objective:To investigate the sensitivity of esophageal squamous cell carcinoma EC9706 cells to rapamycin after silencing mTOR expression by small interfering RNA targeting mTOR (mTOR-siRNA). Methods: EC9706 cells were transfected with mTOR-siRNA and the interference effect was investigated by RT-PCR. EC9706 cells were treated with rapamycin before and after mTOR-siRNA transfection, and the expressions of mTOR and its downstream p70S6K were detected by Western blotting analysis. Cell cycle, apoptosis and proliferation of EC9706 cells were determined by flow cytometry and CCK-8 kit, respectively. Results: mTOR-siRNA down-regulated the expression of mTOR mRNA in EC9706 cells(P<0.05 or P<0.01). Rapamycin inhibited mTOR and phosphorylated p70S6K (p-p70S6K) expressions and increased p70S6K expression in EC9706 cells(all P<0.05), and these effects of rapamycin were further enhanced by mTOR-siRNA transfection (P<0.05). Rapamycin also induced apoptosis, inhibited proliferation and arrested cell cycle in G1 phase of EC9706 cells (all P<0.01), and transfection with mTOR-siRNA significantly promoted these effects of rapamycin in EC9706 cells (P<0.05). Conclusion: mTOR-siRNA can specifically down-regulate mTOR expression in esophageal squamous cell carcinoma EC9706 cells, and increase the sensitivity of EC9706 cells to rapamycin.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 30901778), and the Foundation for Introduced Talents of Zhengzhou University
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