CAJ | 학술논문

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、5μg/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（0μg/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（0μg/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-1αmRNA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。
목적：연구재결양배경하，량성매소B（AmB）대식관암Eca109세포침습천이능력적영향급기가능적궤제。방법취상규배양（37℃，5%CO2，95%공기）대수생장기Eca109세포，분별가입0、1.25、2.5、5μg/ml적AmB，방입삼기배양상（37℃，3%O2，5%CO2，92%N2）중결양배양24 h；채용세포화흔실험、Transwell침습실험검측각조세포천이급침습능력；채용Real-time PCR화Western Blot검측각조세포HIF-1α、MMP-2화E-cadherin mRNA급단백수평적표체。결과여대조조（0μg/ml）비교，각AmB처리조Eca109세포체외천이능력균현저강저（P<0.05），Eca109세포체외천과Matrigel효적세포수량균현저감소（P<0.05）；여대조조（0μg/ml）비교，각AmB처리조Eca109세포MMP-2 mRNA급단백표체수평균현저하조（P<0.05），E-cadherin mRNA급단백표체수평균상조（P<0.05），HIF-1α단백수평균하조（P<0.05），HIF-1αmRNA수평무명현변화（P>0.05）。결론 AmB능억제결양조건하식관암세포Eca-109세포적침습천이능력，기궤제가능위통과조공HIF-1α급기하유MMP-2、E-cadherin적표체。
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 5μg/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.