Analysis of differentially expressed genes in endothelial cells following tumor cell adhesion and role of PRKAA2 and miR-124-3p

Tumor cell adhesion to endothelium is one pattern of tumor-endothelium interaction and one key step during tumor metastasis. The endothelium integrity is an important barrier to prevent tumor invasion and metastasis. Changes happened in ECs according to tumor cells adhesion provide important signaling mechanisms for the angiogenesis and metastasis of tumor cells. However, it is unclarified by now. In this present study we used Affymetrix Gene Chip Human Transcriptome Array 2.0. and quantitative real-time PCR (qPCR) to clarify the detailed genes alteration in endothelial cell adhered by prostate tumor cells PC-3M. A total of 504 differentially expressed mRNAs and 444 lncRNAs were obtained through chip data analysis. Human umbilical vein endothelial cells (HUVECs) were cultured in 96-well dishes coated with 1 % gelatin. When HUVECs reached an approximate confluence of 100 %, PC-3M cells were washed and resuspended in culture medium (10e5 cells/ml). One hundred microliters of the tumor cell suspension were added to HUVECs monolayers in 96-well dishes and co-cultured with HUVECs for 30 min at 37°C. At the end of the incubation, wells were washed three times with PBS to try to remove adherent and non-adherent PC-3M cells.