[The Supernatant Obtained from Cultured Anip973 Cells Enhances the Biological Activities of HUVEC].

Background And Objective: Unlike normal tissue-derived microvascular endothelial cells, tumor microvessel endothelial cells are highly reactive to growth factors and exhibit more adhesion molecules. Thus, vascular tumors are highly permeable and grow vigorously; this occurrence results in rapid growth and metastasis cancer cells. Therefore, understanding the characteristics of endothelial cells in the tumor microenvironment guides anti-angiogenic therapy. To this end, we explore the effect of the supernatant obtained from cultured Anip973 cells (high-metastatic human lung adenocarcinoma cells) on the biological behavior and on the cell surface markers of the human umbilical vein endothelial cell (HUVEC).

Methods: The HUVEC that was cultured in a medium (RPMI-1640 + 10% fetal bovine serum) containing various concentrations of Anip973 supernatants was categorized into experimental groups. The HUVEC cultured in a medium without Anip973 supernatants served as the control group. Proliferation was determined with CCK-8; blood vessel formation was investigated with three-dimensional culture techniques in vitro; and HUVEC migration was observed via transwell assay. At the same time, the expressions of CD105, CD31, and the apoptotic marker of Annexin V were detected through flow cytometry for analyzing the relationship between the expression of cell surface markers and biological behavior.

Results: Following incubation with the supernatant obtained from cultured Anip973 cells, HUVEC proliferated more than the control group did, and the proliferation rate was maximized when incubated in a supernatant concentration of 250 μL/mL for 24 h (P=0.002). In addition, the experimental groups exhibited varying degrees of migration and forms of vascular lumen sample structure, especially at supernatant concentrations of 125 µL/mL (P<0.001) and 250 µL/mL (P=0.002), respectively. CD105 expression was optimized at 250 μL/mL (P=0.028), and CD31 expression also increased with an increase in concentration. However, the percentage of apoptotic cells decreased. Correlation analysis results showed that cell proliferation, migration, and CD105 expression were significantly and positively correlated with one another. By contrast, no significant correlation was detected between CD31 expression and biological behavior.

Conclusions: Anip973 supernatants can promote HUVEC proliferation and migration, as well as angiogenesis. In addition, cell surface markers can change concurrently and relatively. To a certain extent, changes in CD105 expression can be attributed to shifts in its biological behavior.
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