Kang Qiangqiang, Tang Min, Hou Yanli, Duan Liqun, Chen Xingyue, Shu Jin, Wu Furong, Wang Ying, Li Shaolin
Department of Radiology and Oncology, Chongqing Medical University, Chongqing 400016, China.E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2014 Jun;34(6):798-801.
To investigate the effect of amphotericinB (AmB) on migration and invasion of esophageal carcinoma Eca109 cells exposed to hypoxia and explore the molecular mechanisms.
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.
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).
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.
探讨两性霉素B（AmB）对缺氧条件下食管癌Eca109细胞迁移和侵袭的影响，并探究其分子机制。
将常规培养的食管癌Eca109细胞在缺氧条件（3% O₂、5% CO₂和92% N₂）下分别用0、1.25、2.5或5 μg/ml的AmB处理24小时。分别通过细胞划痕试验和Transwell小室实验评估细胞迁移和侵袭能力。采用实时定量PCR和蛋白质印迹法分别检测细胞中缺氧诱导因子-1α（HIF-1α）、基质金属蛋白酶-2（MMP-2）和E-钙黏蛋白的mRNA和蛋白表达。
与对照细胞相比，不同剂量AmB处理的细胞迁移和侵袭能力减弱（P<0.05）。AmB处理导致MMP-2的mRNA和蛋白表达显著降低（P<0.05），E-钙黏蛋白表达增加（P<0.05）；AmB处理后细胞中HIF-1α的蛋白表达显著降低（P<0.05），但其mRNA水平无显著变化（P>0.05）。
AmB可能通过调节HIF-1α、MMP-2和E-钙黏蛋白的表达来抑制缺氧微环境中食管癌Eca109细胞的迁移和侵袭。
