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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。
Objective To investigate the effect of amphotericin B (AmB) on the invasion and migration of esophageal carcinoma Eca109 cells under hypoxia and its possible mechanism. Methods Eca109 cells were cultured in the normal (37 ℃, 5% CO2, 95% air) logarithmic growth phase, and were respectively added into AmB at 0, 1.25, 2.5 and 5 μg / , 5% CO2, 92% N2) for 24 h under hypoxia. Cell migration assay and Transwell invasion assay were used to detect cell migration and invasion. Real-time PCR and Western Blot were used to detect the expression of HIF-1α, MMP-2 and E-cadherin mRNA and protein expression. Results Compared with the control group (0μg / ml), the migration ability of Eca109 cells in each AmB treatment group was significantly decreased (P <0.05), and the number of Eca109 cells in vitro significantly decreased (P <0.05) The mRNA and protein expressions of MMP-2 in Eca109 cells were significantly decreased (P <0.05), the expression of E-cadherin mRNA and protein were all increased in AmB treated group (0μg / ml) (P <0.05), while there was no significant difference in HIF-1α mRNA (P> 0.05). Conclusion AmB can inhibit the invasion and migration of esophageal carcinoma cell line Eca-109 under hypoxia. The mechanism may be through the regulation of the expression of HIF-1α and its downstream MMP-2 and E-cadherin.