Effect of conditioned medium of vascular endothelial cells on the epithelial-mesenchymal transition of hepatocellular carcinoma cells
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摘要: 本文旨在研究间接共培养条件下血管内皮细胞分泌或代谢产生的物质对于肝癌细胞上皮-间质转化（EMT）的影响。体外培养人肝癌细胞系 QGY-7703，并与人脐静脉内皮细胞（HUVEC）条件培养基进行共培养；倒置相差显微镜观察肝癌细胞形态学的变化；划痕实验分析肝癌细胞迁移能力的变化；Western blot 及免疫荧光实验检测条件培养基对肝癌细胞 EMT 相关蛋白表达及分布的影响。结果表明，共培养后，QGY-7703 细胞逐渐由多边形变为纺锤形，迁移能力显著提高，EMT 相关标志物的表达及分布均出现时序性变化。研究结果证实，间接共培养条件下，血管内皮细胞可诱导肝癌细胞发生 EMT。Abstract: This study aims to investigate the effect of substances secreted or metabolized by vascular endothelial cells on epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma cells under indirect co-culture condition. Human hepatocellular carcinoma cell line QGY-7703 was cultured in vitro, and then was co-cultured with conditioned medium of human umbilical vein endothelial cells (HUVEC). The morphological changes of QGY-7703 cells were observed by inverted phase contrast microscopy. The migration ability of QGY-7703 cells was analyzed by scratch-wound assays. The effect of conditioned medium on the expression and distribution of EMT related proteins was detected by Western blot and immunofluorescence assays, respectively. The results showed that the QGY-7703 cells gradually changed from polygonal to spindle shape, the migration ability promoted significantly, and both the expression and distribution of EMT related marker changed in a time-dependent manner after co-culturing. The results confirm that vascular endothelial cells can induce EMT in hepatocellular carcinoma cells under indirect co-culture condition.
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图 1 间接共培养条件下 QGY-7703 细胞形态的变化及长短轴之比的统计图
标尺 = 10 μm；*，P < 0.05
Figure 1. The morphology changes of QGY-7703 cells under indirect co-culture condition and the statistical graph of the ratio of long axis to short axis
scale bar = 10 μm; *, P < 0.05
图 2 间接共培养条件下 QGY-7703 细胞迁移情况的显微镜照片及迁移距离的统计图
标尺 = 500 μm；*，P < 0.05
Figure 2. The microscopy images of QGY-7703 cells migration under indirect co-culture condition and the statistical graph of migration distance
scale bar = 500 μm; *, P < 0.05
图 3 间接共培养条件下 QGY-7703 细胞中上皮标志物 E-cadherin 的表达水平和相对表达量的统计图
*，P < 0.05
Figure 3. The expression level of epithelial marker E-cadherin in QGY-7703 cells under indirect co-culture condition and the statistical graph of relative expression
*, P < 0.05
图 4 间接共培养条件下 QGY-7703 细胞中间充质标志物 N-cadherin 和 Vimentin 的表达水平及相对表达量的统计图*，P < 0.05
Figure 4. The expression level of mesenchymal markers N-cadherin and Vimentin in QGY-7703 cells under indirect co-culture condition and the statistical graph of relative expression
*, P < 0.05
图 5 间接共培养条件下 QGY-7703 细胞中 E-cadherin 和 N-cadherin 分布的变化
绿色荧光：E-cadherin；红色荧光：N-cadherin；标尺 = 20 μm
Figure 5. The changes of distribution of E-cadherin and N-cadherin in QGY-7703 cells under indirect co-culture condition
green: E-cadherin; red: N-cadherin; scale bar = 20 μm
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