肝癌条件培养基对血管内皮细胞形貌和超微结构的影响

Effect of Conditioned Medium for Hepatocellular Carcinoma on Morphology and Ultrastructure of Vascular Endothelial Cells

为观察肝癌细胞(HepG2)条件培养基对人脐静脉血管内皮细胞(HUVEC)形貌和超微结构的影响,建立HepG2细胞条件培养基(HepG2-CM)与HUVEC细胞共培养体系。通过倒置显微镜观察HUVEC细胞形态的变化,通过扫描电镜(SEM)检测HUVEC细胞超微结构的变化,通过原子力显微镜(AFM)分析细胞膜表面粗糙度的变化。对照组细胞之间紧密依靠,细胞膜边缘平整,细胞黏附排列致密;HepG2-CM组细胞之间连接疏散,细胞膜边缘模糊,细胞黏附减弱,两组细胞膜表面高低差(Rp-v)为(454.83±29.99)nm和(501.28±11.78)nm,P=0.028;均方根粗糙度(Rq)为(32.84±4.70)nm和(45.03±3.84)nm,P=0.007;平均粗糙度(Ra)为(25.06±4.09)nm和(34.39±3.58)nm,P=0.014;平均高度(Meant Ht)为(204.03±11.65)nm和(301.48±12.80)nm,P=0;中位数高度(Median Ht)为(206.55±7.46)nm和(313.25±5.89)nm,P=0。结果表明,HepG2-CM能诱导HUVEC细胞黏附减弱,细胞之间连接疏散,“伪足”样突起形成,细胞膜表面粗糙度增加。该研究有助于我们进一步理解肿瘤微环境中血管内皮细胞生物学行为,为抗肿瘤血管生成的研究提供思路。

To observe the effect of hepatoma carcinoma cells(HepG2)conditioned medium on the morphology and ultra-structure of human umbilical vein endothelial cells(HUVEC).The co-culture system of HepG2 conditioned medium(HepG2-CM)and HUVEC was established.The changes of cell morphology were observed by inverted microscope,and the changes of cell ultrastructure were observed by scanning electron microscopy(SEM),then the roughness of cell membrane surface were analyzed by atomic force microscopy(AFM).The cells in control group depended closely on each other,the edges of the cell membrane were flat,and the cell adhesion arrangement was dense.The connections between the cells in HepG2-CM group were evacuated,the edges of the cell membrane were blurred,and cell adhesion was weakened.The peak to valley roughness(Rp-v)of cell membrane surface of the two groups were(454.83±29.99)nm and(501.28±11.78)nm,P=0.028.Root-mean-square roughness(Rq)were(32.84±4.70)nm and(45.03±3.84)nm,P=0.007.Average roughness(Ra)were(25.06±4.09)nm and(34.39±3.58)nm,P=0.014.Meant Ht were(204.03±11.65)nm and(301.48±12.80)nm,P=0.Median Ht were(206.55±7.46)nm and(313.25±5.89)nm,P=0.The results indicated that HepG2-CM induces the weakening of HUVEC cell adhesion,the evacuation of connections between cells,the formation of“pseudopod-like”protrusions,and the increase of roughness of cell membrane surface,which is helpful for us to further understand the biological behavior of vascular endothelial cells in tu-mor microenvironment and provide ideas for anti-tumor angiogenesis.