【摘要】 背景与目的应用cDNA微阵列从食管鳞癌配对组织中得到一批在食管鳞癌中差异表达的基因,其中包括SLP-2(stomatin-likeprotein2,SLP-2),它在食管鳞癌中高表达。为验证SLP-2在食管鳞癌中的高表达,构建SLP鄄2真核表达载体,以探讨SLP鄄2与食管鳞癌发生、发展的关系。方法利用PCR从正常食管上皮cDNA中扩增SLP-2的开放阅读框,将SLP-2重组到pcDNA3.1/myc-His(-)中,构建SLP鄄2的真核表达质粒。然后将重组子导入食管鳞癌细胞系TE12中。半定量RT鄄PCR鉴定转染细胞,获得阳性细胞克隆。采用MTT比色法、平板克隆形成实验、流式细胞术和MTS比色法分析SLP鄄2对TE12细胞的影响。结果SLP鄄2在食管鳞癌中高表达。SLP鄄2基因反义转染抑制TE12细胞SLP鄄2表达时,可抑制细胞生长和增殖,导致S期阻滞,并抑制TE12细胞的粘附功能。结论食管鳞癌中SLP鄄2表达上调促进TE12细胞过度生长和增殖,并参与食管鳞癌的转移。更多还原

【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.更多还原