【摘要】 从基因芯片筛选出差异表达的基因中一个细胞周期蛋白 (cyclin)E2基因 ,深入研究周期蛋白E2在高转移性胃腺癌细胞系RF 4 8细胞中的生物学作用 .首先通过合成硫代磷酸化修饰的反义、正义和错配寡核苷酸片段 ,使用半定量RT PCR和Western印迹方法分别检测被 3种寡核苷酸转染的RF 4 8细胞在 1～ 5d内周期蛋白E2基因及它可能调控下游靶基因之一FGFR基因和蛋白质的表达 ,检测寡核苷酸转染的RF 4 8细胞周期和凋亡细胞 ,观察寡核苷酸转染RF 4 8细胞的软琼脂集落形成、运动能力和体外侵袭能力 .结果表明 ,修饰的反义寡核苷酸在有效地抑制RF 4 8细胞中周期蛋白E2表达上调后 ,RF 4 8细胞的迁徙能力被显著降低 ,其增殖、运动能力没有变化 .周期蛋白E2基因的主要作用并不是促细胞分裂 ,也与细胞的增殖、运动能力无关 ,周期蛋白E2基因的过度表达与胃腺癌的迁徙性存在相关性 ,其触发肿瘤的侵袭性可能通过某种机制调控其下游靶基因之一成纤维细胞生长因子受体 (FGFR)基因的表达来实现的更多还原

【Abstract】 Overexpression of cyclin E2 gene in RF-48 was identified by using cDNA microarray, which suggested that up-regulation of cyclin E2 might contribute to metastatic process of gastric adenocarcinoma. Sense, mismatch, and antisense-oligonucleotide targeting for start codon of cyclin E2 were synthesized. Meanwhile, expression of cyclin E2 and FGFR genes and FGFR protein during 5 days in RF-48 cells were measured by using semi-quantitative RT-PCR and Western-blotting, respectively. The cell cycle and apoptosis of RF-48 cells were assayed by using flow cytometry. Colony formation on soft agar, cell migration and invasion abilities of RF-48 cells were also observed in the three transfected cells. After up-regulation of cyclin E2 in RF-48 cells was effectively inhibited using antisense-oligonucleotide transfection, the invasive ability of RF-48 cells was apparently decreased, with no any apparent difference in its ability of migration and proliferation as compared with controls. The results demonstrated that the major role of cyclin E2 in gastric adenocarcinoma might be associated with the process of invasion, but not cell division, migration and proliferation activity. The invasive ability of RF-48 cells triggered by cyclin E2 might involve in an unknown mechanism that regulates expression of one of its down-stream target genes, FGFR.更多还原