南方医科大学学报

南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (06): 798-.

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两性霉素B通过调节HIF-1α活性抑制缺氧环境下食管癌Eca109细胞的侵袭迁移能力

康强强，唐敏，侯妍利，段丽群，陈兴月，舒锦，吴府容，王颖，李少林   

  • 出版日期:2014-06-20 发布日期:2014-06-20

Amphotericin B suppresses migration and invasion of esophageal carcinoma Eca109 cells
in hypoxic microenvironment by down-regulating hypoxia-inducible factor-1α activity

  • Online:2014-06-20 Published:2014-06-20

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摘要： 目的研究在缺氧环境下，两性霉素B（AmB）对食管癌Eca109细胞侵袭迁移能力的影响及其可能的机制。方法取常规
培养（37 ℃，5% CO2，95%空气）对数生长期Eca109细胞，分别加入0、1.25、2.5、5 μg/ml的AmB，放入三气培养箱（37 ℃，3% O2，
5% CO2，92% N2）中缺氧培养24 h；采用细胞划痕实验、Transwell侵袭实验检测各组细胞迁移及侵袭能力；采用Real-time PCR
和Western Blot检测各组细胞HIF-1α、MMP-2和E-cadherin mRNA及蛋白水平的表达。结果与对照组（0 μg/ml）比较，各AmB
处理组Eca109细胞体外迁移能力均显著降低（P<0.05），Eca109细胞体外穿过Matrigel胶的细胞数量均显著减少（P<0.05）；与对
照组（0 μg/ml）比较，各AmB处理组Eca109细胞MMP-2 mRNA及蛋白表达水平均显著下调（P<0.05），E-cadherin mRNA及蛋
白表达水平均上调（P<0.05），HIF-1α蛋白水平均下调（P<0.05），HIF-1α mRNA水平无明显变化（P>0.05）。结论AmB能抑制缺
氧条件下食管癌细胞Eca-109细胞的侵袭迁移能力，其机制可能为通过调控HIF-1α及其下游MMP-2、E-cadherin的表达。

Abstract: Objective To investigate the effect of amphotericinB (AmB) on migration and invasion of esophageal carcinoma
Eca109 cells exposed to hypoxia and explore the molecular mechanisms. Methods Routinely cultured esophageal carcinoma
Eca109 cells were treated with 0, 1.25, 2.5, or 5 μg/ml AmB in hypoxic condition (3% O2, 5% CO2, and 92% N2) for 24 h. The cell
migration and invasion were assessed by cell scratch test and Transwell chamber assay, respectively. Real-time quantitative
PCR and Western blotting were used to detect the mRNA and protein expressions of hypoxia-inducible factor-1α (HIF-1α),
matrix metalloproteinase-2 (MMP-2), and E-cadherin in the cells, respectively. Results Compared with the control cells, the
cells treated with different doses of AmB showed attenuated ability of migration and invasion (P<0.05). AmB treatment
resulted in significantly lowered mRNA and protein expressions of MMP-2 (P<0.05) and increased expressions of E-cadherin
(P<0.05); the protein expression of HIF-1α decreased significantly in cells after AmB treatment (P<0.05) but its mRNA levels
showed no significant changes (P>0.05). Conclusion AmB can suppress the migration and invasion of esophageal carcinoma
Eca109 cells in hypoxic microenvironment possibly by regulating the expressions of HIF-1α, MMP-2 and E-cadherin.