Malignant phenotype of renal cell carcinoma cells is switched by Ukrain administration in vitro

Abstract

aWe investigated whether Ukrain modulates the malignant phenotype of clear cell renal cell carcinoma (ccRCC) cells Caki-1, Caki-2, and ACHN treated with four doses (5, 10, 20, and 40 lmol/l) for 24 and 48 h. The epithelial-tomesenchymal transition markers E-cadherin, b-catenin, and vimentin were analyzed by immunofluorescence as well as actin and tubulin; matrix metalloproteinase-2 and matrix metalloproteinase-9 activity was analyzed by SDS-zymography, intracellular and secreted SPARC levels by western blot, and cell cycle by flow cytometry. Ukrain did not induce E-cadherin/b-catenin immunoreactivity at the cell-cell boundary, although it determined the actin cortical expression in Caki-2 and ACHN, and did not affect vimentin organization; however, in some Caki-1 and ACHN cells the perinuclear concentration of vimentin was consistent with its downregulation. Matrix metalloproteinase-2 and matrix metalloproteinase-9 activity was significantly downregulated 48 h after 20 lmol/l Ukrain administration. At this time point, Ukrain significantly decreased migration and invasion, and downregulated SPARC levels in cell supernatants at all doses in Caki-2, and at 20 lmol/l in Caki-1 and ACHN cells. Concomitantly, SPARC was upregulated in all ccRCC cells, suggesting that Ukrain could also affect cell proliferation by cell cycle inhibition, as supported by the cell cycle analysis, as SPARC also acts as a cell cycle inhibitor. Our results suggest that Ukrain may switch the epithelial-to-mesenchymal transition-related phenotype of ccRCC cells, and targets the two major aspects involved in RCC progression, such as tumor invasion/microenvironment remodeling and cell proliferation.

ITF promotes migration of intestinal epithelial cells through crosstalk between the ERK and JAK/STAT3 pathways

“…The cell adhesion molecule, E-cadherin, is a transmembrane glycoprotein whose cytoplasmic domain is complexed with β-catenin 15 . According to the literature, several growth factors can induce the loss of E-cadherin expression and function involved in cell adhesion and migration 16 . Victor Y.W.…”

Altered metabolic pathways in clear cell renal cell carcinoma: A meta-analysis and validation study focused on the deregulated genes and their associated networks.

“…The human ccRCC cell lines Caki-2 and ACHN (kindly provided by Dr Nicoletta Gagliano [ 43 ]) were used. Cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 2 mmol/l glutamine, antibiotics (100 U/ml penicillin, 0.1 mg/ml streptomycin) and amphotericin B (2.5 μg/ml).…”