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Archive > Volume 24 Issue 3 > 2017,24(3):247-252. DOI:10.3872/j.issn.1007-385X.2017.03.006 Prev Next
Basic Research
Effect of Bin1 methylation on invasion ability of esophageal squamous cell carcinoma TE13 line cell
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Abstract:
Objective:To detect methylation status of Bin1 in esophageal squamous cell carcinoma (ESCC) TE13 line cell, to analyze changes of expression level of Bin1, its methylation status and bioactivity of the TE13 cell before and after demethylation with demethylation agent 5-Aza-dC, and to explore possible mechanism and therapeutical strategies of ESCC. Methods: Using qRT-PCR, expression of bridge integration factor 1 (Bin1) mRNA in the TE13 cell at pre- and post-demethylation of Bin1 was detected. Methylation status of Bin1 promoter region in the TE13 cells were detected by SDP assay. Effects of demethylation on abilities of migration and invasion of the TE13 cell were respectively tested by scratch and Transwell assays. To use Western blotting assay, expressions of Bin1, matrix metalloproteinases-2 (MMP-2) and MMP-9 proteins in the TE13 cell were detected. Results: Bin1in the TE13 cell was presented as a complete methylation. Expression of Bin1 mRNA was low, and after the treatment of 5-Aza-dC, expression of Bin1 mRNA was significantly increased (P<0.01). Results of scratch and Transwell assays showed that treatment of demethylation evidently decreased migration and invasion abilities of the TE13 cell (all P<0.01). After the treatment with 5-Aza-dC, expression of Bin1 protein was markedly increased, expressions of MMP-2 and MMP-9 proteins were significantly decreased (all P<0.01). Conclusion: DNA methylation could be one of important mechanisms for low expression or absence of Bin1, and the methylation could affected migration and invasion abilities of the TE13 cell by regulating expressions of MMP-2 and MMP-9 proteins.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No.81201607), the Outstanding Youth Foundation of Hebei Province (H2014206320), and the Fundation for High Level Talents of Hebei Province (A201401040)
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