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目的克隆生长抑素受体SSTR3基因,构建其真核表达载体,并进行转染研究,探讨SSTR3基因在胃癌细胞中的作用。方法从永生化的胃上皮细胞系GES中提取总RNA,经RTPCR扩增出SSTR3基因,并构建真核表达载体pcDNA3.1(+)SSTR3,转染胃癌细胞系MKN45,以MTT法和流式细胞仪观察转染SSTR3基因前后使用奥曲肽对胃癌细胞增殖和凋亡的影响。结果克隆的SSTR3基因测序正确;构建的真核表达载体pcDNA3.1(+)SSTR3能高表达SSTR3蛋白;转染SSTR3基因的胃癌细胞系MKN45加用奥曲肽后,出现明显的增殖抑制和凋亡发生。结论SSSTR3基因介导了生长抑素类似物引起的胃癌细胞的增殖抑制和凋亡发生。通过基因转移使原来不表达SSTR3基因的胃癌细胞再表达,为生长抑素和SSTR3基因治疗胃癌提供了又一有效途径。
Objective To clone the somatostatin receptor SSTR3 gene and construct its eukaryotic expression vector and transfect it to investigate the role of SSTR3 in gastric cancer cells. Methods The total RNA was extracted from the immortalized gastric epithelial cell line GES. The SSTR3 gene was amplified by RTPCR. The eukaryotic expression vector pcDNA3.1 (+) SSTR3 was constructed and transfected into gastric cancer cell line MKN45. MTT assay and flow cytometry Effects of octreotide on proliferation and apoptosis of gastric cancer cells before and after SSTR3 gene transfection were observed by cytometry. Results The cloned SSTR3 gene was sequenced correctly. The constructed eukaryotic expression vector pcDNA3.1 (+) SSTR3 could highly express SSTR3 protein. After transfected with SSTR3 gene, gastric cancer cell line MKN45 plus octreotide showed obvious proliferation inhibition and apoptosis . Conclusion SSSTR3 gene mediates somatostatin analogue-induced gastric cancer cell proliferation and apoptosis. The re-expression of gastric cancer cells that originally did not express SSTR3 gene by gene transfer provides yet another effective approach for the treatment of gastric cancer by somatostatin and SSTR3 gene.