Study of miR-29c inhibiting invasion and migration of ESCC cell by targeting Tiam1

doi:10.3969/j.issn.1673-8640.2015.06.022

Abstract

Abstract: Objective

To research microRNA(miR)-29c inhibiting invasion and migration of esophageal squamous cell carcinoma(ESCC) cell by targeting T-lymphom invasion and metastasis 1(Tiam1).

Methods

Invasion and migration capacity changes of ESCC cell with overexpression of miR-29c were observed through cell scratch and transwell experiments. Western blot analysis and sensor reporter assay were used to research the regulation and target site of miR-29c for Tiam1. In shRNA rescue experiments, Western blot analysis was used to detect the expression change. Immunofluorescence-laser scanning confocal microscope method was used to investigate the mechanisms of miR-29c regulating the invasion and migration capacities of ESCC cell.

Results

Cell scratch and transwell experiment results showed that miR-29c inhibited the invasion and migration of ESCC cell. Tiam1 was predicted to be the target gene of miR-29c through bioinformatics methods. Western blot analysis and sensor reporter assay showed that miR-29c degradated Tiam1 through binding Tiam1 3'-untranslated region (UTR). The analysis of shRNA rescue experiments, including cell scratch experiment, Western blot and immunofluorescence analysis, showed that activated Rac1- guanosine triphosphate(GTP) reduced due to miR-29c inhibiting Tiam1 expression, which resulting migration capacity of ESCC cell diminished, while the inter ference of siTiam1 (artificially synthesized siRNA-specific molecule for Tiam1) can "rescue" the migration capacity of ESCC cell.

Conclusions

miR-29c lowers the level of Rac1 activation through degradating Tiam1 mRNA, resulting the change of cell motility status, thereby inhibiting ESCC cell invasion and migration.

Key words: MicroRNA-29c, T-lymphom invasion and metastasis 1, Esophageal squamous cell carcinoma, Invasion and migration

CLC Number:

R393

CHU Qinghua, ZHANG Jie, SUN Fenyong. Study of miR-29c inhibiting invasion and migration of ESCC cell by targeting Tiam1[J]. Laboratory Medicine, 2015, 30(6): 635-640.

Figures/Tables 7

References 19

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