CLONING AND EXPRESSION OF TYRO3 ISOFORMS IN BLADDER CANCER CELLS

Abstract

Bladder cancer is the most prevalent cancers diagnosed worldwide with poor prognoses and low survival rates. Recent attention has been paid to the TAM family receptors, including TYRO3, AXL, and MERTK which are key oncogenes in several malignancies. Previous study has investigated the high expression of TYRO3 in bladder cancer cells. However, little is known about TYRO3 Isoforms and their subcellular localization in bladder cancer cells. To our knowledge, TYRO3 Isoform I is a full-length receptor, whereas Isoform II doesn’t contain exon 1 which is in charge of encoding the signal peptide. It was hypothesized that TYRO3 Isoforms are present in the nucleus. In the present work, plasmids encoding TYRO3 Isoform I and II fused with HA tag were obtained via molecular cloning. Gene sequencing has confirmed the similarity with TYRO3 transcript variants via BLASTn tool. Recombinant plasmids were transfected into T24 and UMUC3 bladder cancer cells and protein expression was detected. The subcellular localization of TYRO3 transcript variants were determined. Interestingly, TYRO3 Isoform I was present in the membrane and localized in the spindle positioning. TYRO3 Isoform II, on the other hand, was present in the perinuclear region. Therefore, the study highlights the relevance of transcript variants in bladder cancer.