Expression of carbonic anhydrase III (CAIII) from different cell lines
Abstract
Carbonic anhydrase III (CAIII, Carbonate Dehydratase EC 4.2.1)is an abundant cytoplasmic protein found in skeletal muscle, liver,and adipocytes. It is distinguished from other members of the CAfamily by characteristics such as very low carbon dioxide hydra-tase activity, resistance to sulfonamide inhibitors, and resistanceto oxidant-mediated S-glutathiolation. In addition CAIII is acharacteristic of slow-twitch, aerobic, type I muscle fibers. Somestudies with CAIII showed that CAIII is involved in mitochon-drial ATP synthesis indicating a role of CAIII in cystic fibrosisliver disease. CAIII, is considered as one of the proteins involvedin oxidative stress response both in liver and skeletal muscle,probably acting as a scavenger of reactive oxygen species (ROS)and, thus protecting cells from oxidative damage. CAIII seems toplay an important role also in disruption of the intercellular bar-rier associated with the down-regulation of E-cadherin in the lar-yngopharyngeal reflux disease. In recent years, CAIII relationshipwith the cancer was also investigated. The contributions of inva-sion have been demonstrated in the early stages of metastasis.There is limited information about the expression pattern ofCAIII. In our study, we evaluated the expression of CAIIImRNA expression profile in different cell lines, namely, HumanUmbilical Vein Endothelial Cells (HUVEC), Human osteosar-coma cell line(MG63), Human epithelial-like osteosarcoma cellline (Saos-2) and Human epithelial-like prostate carcinoma cellline (DU145). For this purpose cells were cultured with DMEMHigh Modified Medium, 10% fetal calf serum solution. Cell via-bility controlled by trypan blue exclusion. Total cellular RNAwas prepared and subjected to RT-PCR using CAIII and internalcontrol GAPDH specific primers. Amplified fragments were ana-lyzed by agarose gel electrophoresis. CAIII expression is detectedin HUVEC, MG63 and Saos-2 cell lines. However, CAIII is notexpressed in DU145 cell line.