Ectopic expression of TWIST1 upregulates the stemness marker OCT4 in the esophageal squamous cell carcinoma cell line KYSE30.

BACKGROUND

The transcription factor TWIST1 plays an important role in the epithelial-mesenchymal transition (EMT) process and in the migration, invasion and metastasis of cancer cells. OCT4, which is a homeobox transcription factor, has an important role in the self-renewal potential of cancer cells. Our aim here is to elucidate impact of ectopic expression of TWIST1 on OCT4 gene expression in esophageal squamous cell carcinoma (ESCC).

METHODS

The ESCC line was KYSE30. GP293T cells were transfected with purf-IRES-GFP and pGP plasmids to produce recombinant viral particles. A semi-confluent KYSE30 culture was transduced with the prepared retroviral particles. mRNA extraction and cDNA synthesis were performed from normal KYSE30 cells and those ectopically expressing . Expressional analysis of and were performed with relative comparative real-time PCR.

RESULTS

Ectopic expression of in KYSE30 cells was related to its significant overexpression: nearly nine-fold higher in GFP-h KYSE-30 cells than in control GFP cells. This induced expression of caused significant upregulation of in GFP-h KYSE-30 cells: nearly eight-fold higher. In silico analysis predicted the correlation of and through .

CONCLUSIONS

Overexpressed can be correlated with upregulation of the cancer stem cell marker and the protein may play critical regulatory role in gene expression. Since OCT4 is involved in the self-renewal process, the results may suggest a new linkage between and in the cell biology of ESCC, highlighting the probable role of TWIST1 in inducing self-renewal.