论文部分内容阅读
目的:探讨缺氧环境下缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)、β-连环蛋白(β-catenin)、胰岛素样生长因子ⅡmRNA结合蛋白3(insulin-like growth factorⅡmRNA-binding protein3,IMP3)和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)在胃癌SGC-7901细胞增殖中的作用。方法:将pcDNA6.2-GW/EmGFP-miR-β-catenin质粒转染至SGC-7901细胞,建立稳定转染细胞株miR-β-catenin-7901。实验分成对照组(SGC-7901细胞在常氧条件下培养)、缺氧组(SGC-7901细胞在缺氧条件下培养)、干扰组(miR-β-catenin-7901细胞在常氧条件下培养)和缺氧干扰组(miR-β-catenin-7901细胞在缺氧条件下培养),采用平板克隆形成实验和细胞倍增时间检测各组SGC-7901细胞的增殖情况,蛋白质印迹法检测各组SGC-7901细胞中HIF-1α、β-连环蛋白、IMP3和PCNA蛋白的表达水平。结果:与对照组比较,缺氧组细胞的平板克隆形成数增多、倍增时间缩短(P<0.05);与缺氧组比较,缺氧干扰组细胞平板克隆形成数减少、倍增时间延长(P<0.05)。缺氧组细胞中HIF-1α、β-连环蛋白、IMP3和PCNA蛋白表达水平均高于对照组(P<0.05);干扰组细胞中HIF-1α、β-连环蛋白、IMP3和PCNA蛋白表达水平低于对照组(P<0.05);缺氧干扰组细胞中HIF-1α、β-连环蛋白、IMP3和PCNA蛋白表达水平低于缺氧组(P<0.05)。结论:胃癌细胞的增殖可能与HIF-1α和β-连环蛋白相互作用而上调IMP3和PCNA的表达水平有关。
Objective: To investigate the effects of hypoxia-inducible factor-1α (HIF-1α), β-catenin, insulin-like growth factor Ⅱ mRNA -binding protein3, IMP3) and proliferating cell nuclear antigen (PCNA) in the proliferation of gastric cancer SGC-7901 cells. Methods: The pcDNA6.2-GW / EmGFP-miR-β-catenin plasmid was transfected into SGC-7901 cells to establish a stable transfected cell line miR-β-catenin-7901. The experiment was divided into control group (SGC-7901 cells cultured in normoxia), hypoxia group (SGC-7901 cells cultured in hypoxia), interference group (miR-β-catenin-7901 cells cultured in normoxia ) And hypoxia interference group (miR-β-catenin-7901 cells were cultured in hypoxia). The proliferation of SGC-7901 cells was detected by plate clone formation assay and cell doubling time. The expression of SGC-7901 -7901 cells HIF-1α, β-catenin, IMP3 and PCNA protein expression levels. Results: Compared with the control group, the number of plate clone formation and the doubling time of hypoxia group were shorter than those of the control group (P <0.05). Compared with the hypoxia group, the number of plate clone formation and the doubling time prolonged in hypoxia group (P < 0.05). The expression levels of HIF-1α, β-catenin, IMP3 and PCNA in hypoxia group were higher than those in control group (P <0.05). The expression of HIF-1α, β-catenin, (P <0.05). The expression of HIF-1α, β-catenin, IMP3 and PCNA in hypoxia group was lower than that in hypoxia group (P <0.05). Conclusion: The proliferation of gastric cancer cells may be related to the interaction between HIF-1α and β-catenin and the up-regulation of the expression of IMP3 and PCNA.