[Effect of stomatin-like protein 2 (SLP-2) gene on growth and proliferation of esophageal squamous carcinoma cell line TE12]

Abstract

Background & objective: Stomatin-like protein 2 (SLP-2), a differentially expressed gene obtained from esophageal squamous cell carcinoma (ESCC) matched tissues using cDNA microarray, is over-expressed in ESCC tissues. This study was to confirm over-expression of SLP-2 in ESCC tissues, to construct eukaryotic expression plasmid of SLP-2, and investigate the role of up-regulation of SLP-2 in initiation and progression of ESCC.

Methods: Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect expression of SLP-2 in ESCC tissues. Full-length open-reading frame (ORF) of SLP-2 was amplified from cDNA of normal esophageal epithelia by PCR, and inserted into pcDNA3.1/myc-His(-) to construct a eukaryotic expression plasmid of SLP-2. Then the recombinant was transfected into ESCC cell line TE12. Positive cell clones were selected by semi-quantitative RT-PCR. MTT assay, plate clone formation assay, flow cytometry, and MTS assay were performed to measure the effect of SLP-2 on TE12 cells.

Results: SLP-2 was over-expressed in ESCC tissues. Sense/antisense eukaryotic expression plasmids of SLP-2 were constructed. Antisense transfection of SLP-2 gene led to S phase arrest, decreased expression of SLP-2 in TE12 cells, and suppressed cell growth and proliferation, and cell adhesive ability.

Conclusion: Up-regulation of SLP-2 gene might contribute to hyperproliferation of TE12 cells, and metastasis of ESCC.