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AIM To explore the effect of mi R-382 on esophageal squamous cell carcinoma(ESCC) in vitro and its possible molecular mechanism.METHODS Eca109 cells derived from human ESCC and Het-1A cells derived from human normal esophageal epithelium were used. Lentivirus-mediated mi R-382 was overexpressed in Eca109 cells. The effect of mi R-382 on cell proliferation was evaluated by MTT and colony formation assay. For cell cycle analysis, cells were fixed and stained for 30 min with propidium iodide(PI) staining buffer containing 10 mg/m L PI and 100 mg/m L RNase A, and analyzed by BD FACSCalibur? flow cytometer. For cell apoptosis assay, cells were stained with an Annexin V-FITC/PI Apoptosis Detection Kit according to the manufacturer’s instructions and analyzed by a dual-laser flow cytometer. Cell invasion and migration abilities were determined through use of transwell chambers, non-coated or pre-coated with matrigel. Levels of proteins related to cell growth and migration were examined by western blotting.RESULTS Endogenous mi R-382 was down-regulated in Eca109 cells compared with Het-1A. Introduction of mi R-382 not only significantly inhibited proliferation and colony formation, but also arrested cell cycle at the G2/M phase, as well as promoted apoptosis and autophagy in Eca109 cells. Migration, invasion and epithelialmesenchymal transition of Eca109 cells were suppressed by overexpressing mi R-382. Western blotting results showed that mi R-382 inhibited the phosphorylation of m TOR and 4E-BP1. CONCLUSION mi R-382 functions as a tumor suppressor against ESCC development and metastasis, and could be considered as a potential drug source for the treatment of ESCC patients.
AIM To explore the effect of mi R-382 on esophageal squamous cell carcinoma (ESCC) in vitro and its possible molecular mechanism. METHODS Eca109 cells derived from human ESCC and Het-1A cells derived from human normal esophageal epithelium were used. Lentivirus-mediated The effect of mi R-382 on cell proliferation was evaluated by MTT and colony formation assay. For cell cycle analysis, cells were fixed and stained for 30 min with propidium iodide (PI) staining buffer containing 10 mg / ml PI and 100 mg / ml RNase A, and analyzed by BD FACSCalibur® flow cytometer. For cell apoptosis assay, cells were stained with an Annexin V-FITC / PI Apoptosis Detection Kit according to the manufacturer’s instructions and analyzed by a dual-laser flow cytometer. Cell invasion and migration abilities were determined through use of transwell chambers, non-coated or pre-coated with matrigel. Levels of proteins related to cell growth and migration were examined by wes tern blotting .RESULTS Endogenous mi R-382 was down-regulated in Eca109 cells compared with Het-1A. Introduction of mi R-382 not only significantly inhibited proliferation and colony formation, but also arrested cell cycle at the G2 / M phase, as well as Popular apoptosis and autophagy in Eca109 cells. Migration, invasion and epithelialmesenchymal transition of Eca109 cells were suppressed by overexpressing mi R-382. Western blotting results showed that mi R-382 inhibited the phosphorylation of m TOR and 4E-BP1. CONCLUSION mi R-382 functions as a tumor suppressor against ESCC development and metastasis, and could be considered as a potential drug source for the treatment of ESCC patients.