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


CONCLUSIONS
Up-regulation of SLP-2 gene might contribute to hyperproliferation of TE12 cells, and metastasis of ESCC.
Zhang
Li-Yong
LY
MOE Key Laboratory of Bioinformatics, Department of Automation, Tsinghua University, Beijing, 100084, P. R. China.
Ding
Fang
F
Liu
Zhong-Min
ZM
Li
Wen-Dong
WD
Liu
Zhi-Hua
ZH
Li
Yan-Da
YD
D001798:Blood Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D002294:Carcinoma, Squamous Cell / Q000378:metabolism / Q000473:pathology*; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D016373:DNA, Antisense / Q000235:genetics*; D004938:Esophageal Neoplasms / Q000378:metabolism / Q000473:pathology*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D016366:Open Reading Frames; D010957:Plasmids; D016196:S Phase; D014162:Transfection
D001798:Blood Proteins; D016373:DNA, Antisense; D008565:Membrane Proteins; C406287:STOML2 protein, human