| 摘要: |
| 目的 研究模拟低氧条件对舌鳞癌SCC-15细胞增殖、周期、凋亡的影响，及SOX2和OCT4蛋白、mRNA的表达变化。方法 体外培养舌鳞癌SCC-15细胞，加入模拟低氧物去铁胺（desferrioxamine，DFO）模拟低氧环境，为模拟低氧组，同时设置不加DFO的常氧组为对照，采用CCK-8法检测SCC-15细胞的增殖率；流式细胞技术检测SCC-15细胞的周期和细胞凋亡率；蛋白质印迹法检测常氧组和模拟低氧组SCC-15细胞中低氧诱导因子1α（HIF-1α）、SOX2及OCT4蛋白的表达；qPCR检测常氧组和模拟低氧组SCC-15细胞中HIF-1α、SOX2及OCT4 mRNA的表达。结果 与常氧组相比，模拟低氧组SCC-15细胞的增殖受到抑制（P<0.05），G1期细胞所占比例上升、S+G2期细胞所占比例下降（P<0.05），细胞凋亡率未见明显变化（P>0.05）；蛋白质印迹法结果显示，与常氧组比较，模拟低氧组SCC-15细胞中HIF-1α、SOX2、OCT4蛋白的表达均升高，且HIF-1α、OCT4蛋白的升高幅度大于SOX2蛋白，差异有统计学意义（P<0.05，P<0.01）；qPCR结果显示，两组SCC-15细胞中HIF-1α mRNA水平差异无统计学意义（P>0.05），而模拟低氧组SCC-15细胞中SOX2、OCT4 mRNA的表达水平高于常氧组（P<0.05）。结论 DFO可以有效模拟低氧环境，促进SOX2和OCT4的蛋白和mRNA表达水平升高。 |
| 关键词: 舌肿瘤 细胞低氧 去铁胺 SCC-15细胞 肿瘤干细胞 转录因子 |
| DOI：10.16781/j.0258-879x.2017.07.0891 |
| 投稿时间：2016-12-05修订日期：2017-02-27 |
| 基金项目:上海市浦东新区卫生和计划生育委员会青年科技基金项目（PW2014B-24），第二军医大学附属公利医院院级优秀青年基金项目资助（GL2014Q-19）. |
|
| Effect of simulated hypoxia on expression SOX2 and OCT4 of tongue squamous cell carcinoma SCC-15 cells |
| WANG Kai*,JING De-bao,YU Su-ping,TANG Wei,LIU Xue-yang |
(Department of Stomatology, Gongli Hospital, Second Military Medical University, Shanghai 200135, China
*Corresponding author) |
| Abstract: |
| Objective To explore the effect of simulated hypoxic environment on proliferation, cell cycle, apoptosis and the protein and mRNA expressions of SOX2 and OCT4 in tongue squamous cell carcinoma SCC-15 cells. Methods SCC-15 cells was cultured in vitro, and the hypoxia mimetic agent desferrioxamine (DFO) was added to simulate hypoxic condition (simulated hypoxia group, HOX group) and normoxia was designed as control group (NOX group). CCK-8 assay was performed to measure the proliferation ability of the SCC-15 cells; flow cytometry was used to measure the apoptosis rate and cell cycle of the SCC-15 cells; Western blotting was applied to measure the expressions of HIF-1α, SOX2 and OCT4 protein in NOX and HOX groups; qPCR was used to measure the expressions of HIF-1α, SOX2 and OCT4 mRNA in NOX and HOX groups. Results DFO effectively simulated the hypoxic condition. Compared with the NOX group, the proliferation ability of the cells in HOX group was significantly inhibited by DFO (P<0.05), the proportion in G1 cells was increased and that in S+G2 was decreased; while no significant difference was found in the apoptosis rate of SCC-15 cells; the expressions of HIF-1α, SOX2 and OCT4 proteins were significantly increased in HOX group, and the expressions of HIF-1α and OCT4 proteins were significantly higher than SOX2 (P<0.05). There was no significant difference of mRNA expression of HIF-1α in NOX and HOX groups (P>0.05); the mRNA expressions of SOX2 and OCT4 in HOX group were significantly higher than those in NOX group (P<0.05). Conclusion DFO can effectively simulate hypoxic environment, and hypoxia can promote the expressions of SOX2 and OCT4 proteins and mRNA. |
| Key words: tongue neoplasms cell hypoxia deferoxamine SCC-15 cells neoplastic stem cells transcription factors |
