摘要
目的探讨糖原分支酶1(GBE1)在缺氧肺癌组织和细胞中的表达情况,评估其对肺癌缺氧耐受的意义,并探讨其影响肺癌凋亡的机制。方法通过GEO数据库下载含20例缺氧肺癌和常氧肺癌组织基因芯片的数据,结合GCBI平台,筛选差异基因进行GO和KEGG通路分析;体外培养A549细胞,敲降或高表达GBE1,缺氧处理后,MTT法检测细胞活力,Western blot和Real-time PCR检测凋亡相关基因的表达。结果 20例肺癌标本中,差异基因共206个,GBE1为差异最显著,GO及KEGG通路分析后发现差异基因涉及细胞周期、细胞代谢等多个通路;A549细胞缺氧处理后GBE1表达水平显著升高,敲降GBE1能显著增强A549细胞缺氧处理后细胞凋亡相关分子的表达,高表达GBE1能够抑制细胞凋亡相关分子的表达。结论 GBE1可能是潜在的造成肺癌缺氧耐受的分子标志物,为进一步研究肺癌的缺氧耐受机制提供参考。
Objective To investigate the expression of glucogen branching enzyme 1 （ GBE1 ） in hypoxic lung cancer tissues and cells,to evaluate its significance for hypoxia tolerance in lung cancer and to explore its mecha- nism of apoptosis of lung cancer. Methods Data of 20 patients with hypoxic lung cancer and normoxia lung cancer tissue microarray were downloaded through the GEO database, GCBI was used to screen the differential genes for GO and KEGG pathway analysis. In vitro culture of A549 cells was cultured and transfected with siGEB1 for knock down or GBE1 for overexpression. After hypoxia treatment, cell viability was analyzed using MTT assay. Western blot and real-time PCR were used to detect the expression of apoptosis-related genes. Results The data from 20 cases of lung cancer specimens showed that 206 genes displayed differences, and the GBE1 was the most signifi- cant, GO and KEGG pathway analysis found that the differential gene involved in cell cycle, cell metabolism and other multiple access. The expression of GBE1 in A549 cells after hypoxia treatment was significantly increased, and GBE1 could significantly enhance the expression of apoptosis-related genes in A549 cells after hypoxia treat- ment. Over-expression of GBE1 could inhibit the expression of apoptosis-related genes. Conclusion GBE1 may be a potential marker of hypoxia tolerance in lung cancer, this study provide a reference for further study of hypoxia tol- erance mechanism of lung cancer.
作者
丁翔
葛圣林
张朝东
Ding Xiang1,2, Ge Shenglin1 , Zhang Chaodong2(1Dept of Thoracic Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei 230022 ; 2Dept of Thoracic Surgery,Anhui Provincial Chest Hospital,Hefei 23003)
出处
《安徽医科大学学报》
CAS
北大核心
2018年第3期359-364,共6页
Acta Universitatis Medicinalis Anhui
基金
安徽省自然科学基金(编号:KJ2013Z108)
