Qiu Hong, Hu Guangyuan, Xiong Huihua
Oncology Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2011 Feb;31(1):52-57. doi: 10.1007/s11596-011-0149-1. Epub 2011 Feb 19.
The chronic-hypoxia-resistant gastric cancer cell line was established, and its biological characteristics were explored and compared with the parental cell line. Gastric cancer cell lines were cultured under the degressive oxygen concentration. Cell doubling time was calculated by cell counting method. Chemo-resistance ability of cells was tested by MTT assay. Irradiation tolerance of cells was evaluated by colony forming method. Cell cycle distribution was tested with flow cytometry. Invasive ability was tested by Transwell method. The expression levels of GLUT-1 and HIF-1α were detected by using Western blot. MNK45/HYP cells successfully survived under the 1% concentration of oxygen and its cell doubling time was 35.01±1.02 h, while that of MNK45 was 27.35±0.83 h (P<0.01). The percentage of MNK45/HYP cells in G(0)/G(1) stage was (58.3±6.1)%, and that of MNK45 cells was (42.2±6.0)% (P<0.05). Comparing with the parental cells MNK45, drug resistance indexes of 5-Fu, PTX, OXA, Sn38, GEM and VP16 in MNK45/HYP cells were respectively 5.3, 1.3, 3.6, 2.2, 4.8 and 4.4. Colony forming ability of MNK45/HYP cells after irradiation was also significantly higher than MNK45 cells. The invasive number of MNK45/HYP cells was 107.7±17.5, while that of MNK45 cells was 59.0±9.9. The expression levels of GLUT-1 and HIF-1α in MNK45/HYP cells were significantly higher than those in MNK45 cells. MNK45/HYP cells hold biological characteristics of hypoxia tumor with good tolerance to chronic hypoxia, and can be used for the research of solid tumor under chronic hypoxia condition.
建立了耐慢性低氧胃癌细胞系，并探究了其生物学特性，并与亲代细胞系进行比较。胃癌细胞系在递减的氧浓度下培养。通过细胞计数法计算细胞倍增时间。通过MTT法检测细胞的化疗耐药能力。通过集落形成法评估细胞的辐射耐受性。用流式细胞术检测细胞周期分布。通过Transwell法检测侵袭能力。使用蛋白质免疫印迹法检测GLUT-1和HIF-1α的表达水平。MNK45/HYP细胞在1%氧浓度下成功存活，其细胞倍增时间为35.01±1.02小时，而MNK45细胞的细胞倍增时间为27.35±0.83小时（P<0.01）。MNK45/HYP细胞在G(0)/G(1)期的比例为(58.3±6.1)%，而MNK45细胞的比例为(42.2±6.0)%（P<0.05）。与亲代细胞MNK45相比，MNK45/HYP细胞对5-Fu、PTX、OXA、Sn38、GEM和VP16的耐药指数分别为5.3、1.3、3.6、2.2、4.8和4.4。MNK45/HYP细胞照射后的集落形成能力也显著高于MNK45细胞。MNK45/HYP细胞的侵袭数为107.7±17.5，而MNK45细胞的侵袭数为59.0±9.9。MNK45/HYP细胞中GLUT-1和HIF-1α的表达水平显著高于MNK45细胞。MNK45/HYP细胞具有低氧肿瘤的生物学特性，对慢性低氧具有良好的耐受性，可用于慢性低氧条件下实体瘤的研究。
