Down-regulation of TWIST decreases migration and invasion of laryngeal carcinoma Hep-2 cells by regulating the E-cadherin, N-cadherin expression

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Abstract

Purpose

The transcription factor TWIST is an important factor in regulation of the epithelial–mesenchymal transition (EMT), which represents the primary stages during the metastasis of tumors. To identify the role of TWIST in the regulation of metastasis in laryngeal carcinoma Hep-2 cells, we investigated whether the alteration of TWIST has an effect on the Hep-2 cells morphology and whether the alteration of TWIST has an effect on the expression of E-cadherin, N-cadherin as well as the ability of cell motion, migration, and invasion.

Methods

Morphological changes of Hep-2 cells that were transfected a mircoRNA against TWIST vector were observed by the reserved microscope. Reverse transcription-polymerase chain reaction was performed in order to examine the mRNA expression of TWIST, E-cadherin, and N-cadherin. Western blotting was performed to examine the protein expression of TWIST, E-cadherin, and N-cadherin. Cell motion ability was examined by Scratch-wound assay. Transwell™ chamber assays were used to determine cell migration and invasion.

Results

Transfecting a mircoRNA down-regulated TWIST expression at mRNA and protein levels. Down-regulation of TWIST expression induced morphological changes, such as the inversion of the EMT. Moreover, down-regulation of TWIST expression up-regulated E-cadherin and down-regulated N-cadherin expressions at mRNA and protein levels, respectively. Furthermore, we confirmed that down-regulation of TWIST expression decreased the motion, invasion, and migration ability of the Hep-2 cells.

Conclusions

Down-regulation of TWIST expression decreases migration and invasion of laryngeal carcinoma Hep-2 cells by regulation of the E-cadherin, N-cadherin expression.

Keywords

TWIST Laryngeal carcinoma Hep-2 cells Metastasis Epithelial–mesenchymal transition (EMT) 

Abbreviations

EMT

Epithelial–mesenchymal transition

HNSCC

Head and neck squamous cell carcinoma

DMEM

Dulbecco modified eagle medium

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Notes

Acknowledgments

This work was supported in part Department of Science & Technology of Shandong province (03BS020) and by Shandong Province Outstanding Young Scientist Research Award Fund (bs2009yy018).

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Department of Otolaryngology-Head and Neck SurgeryProvincial Hospital Affiliated to Shandong UniversityJinanPeoples’ Republic of China
  2. 2.Department of Otolaryngology-Head and Neck SurgeryDalian Municipal Friendship HospitalDalianPeoples’ Republic of China
  3. 3.Institute of Eye and Otolaryngology, Shandong Clinic Research InstituteJinanPeoples’ Republic of China

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