Silencing of carbonic anhydrase 9 and tetraspanin-8 caused decrease at invasion capacity of human pancreatic carcinoma (PANC-1) cells

Abstract

Carbonic anhydrases (CAs) (EC 4.2.1.1) are zinc-containing met-alloenzymes that catalyze the hydration of CO2molecule anddehydration of HCO3-ions. Carbonic anhydrase 9 is expressed inmany solid tumors and plays a significant role in tumor acid-base homeostasis. Depletion of CA9 gene expression or inhibition ofits catalytic activity shown to retard tumor growth in murinemodels and reduce metastasis.Tetraspanins are integral mem-brane proteins playing a role as organizers of multimolecularcomplexes in the plasma membrane. The human tumor-associ-ated antigen CO-029 (TSPAN8) is a monoclonal antibody-defined cell surface glycoprotein and described as metastasis-pro-moting in several tumor systems. In this study, we aimed to eval-uate the effects of silencing genes, CA9, and TSPAN8, on theinvasion properties of human pancreatic carcinoma (Panc1) cells.Therefore, cells were transfected with specific siRNAs with siR-NA transfection method along with control siRNAs for non-spe-cific targets. Upon silencing, mRNA and protein assays showthat the expression of CAIX and TSPAN8 were decreased com-pared to control cells. siRNA transfected cells were subjected tocell cytotoxicity assay at different time points in order to see ifthe silencing result in an proliferative effect. In addition, the met-astatic and proliferative profiles of CAIX and TSPAN8 weredetermined after in order to 72 and 92 hours by matrigel and clo-nogenic assay.