RRC ID 63857
Author Puteri MU, Watanabe Y, Wardhani BWK, Amalia R, Abdelaziz M, Kato M.
Title PMEPA1/TMEPAI isoforms function via its PY and Smad-interaction motifs for tumorigenic activities of breast cancer cells.
Journal Genes Cells
Abstract PMEPA1 (prostate transmembrane protein, androgen-induced 1)/TMEPAI (transmembrane prostate androgen-induced protein) is highly expressed in diverse cancers, including breast, lung and prostate cancers. It consists of four isoforms with distinct extracellular regions (isoforms a-d). The expression and function of these isoforms are still poorly understood. Hence, we aimed to identify the preferentially expressed isoforms in breast cancer cells and analyze possible differences in tumorigenic functions. In this study, we used 5' Rapid Amplification of cDNA Ends (RACE) and Western blot analyses to identify the mRNA variants and protein isoforms of TMEPAI and found that TMEPAI isoform d as the major isoform expressed by TGF-β stimulation in breast cancer cells. We then generated CRISPR/Cas9-mediated TMEPAI knockout (KO) breast cancer cell lines and used a lentiviral expression system to complement each isoform individually. Although there were no clear functional differences between isoforms, double PPxY (PY) motifs and a Smad-interaction motif (SIM) of TMEPAI were both essential for colony and sphere formation. Collectively, our results provide a novel insight into TMEPAI isoforms in breast cancer cells and showed that coordination between double PY motifs and a SIM of TMEPAI are essential for colony and sphere formation but not for monolayer cell proliferation.
Volume 25(6)
Pages 375-390
Published 2020-6-1
DOI 10.1111/gtc.12766
PMID 32181976
MeSH Amino Acid Motifs Animals Breast Neoplasms / genetics Breast Neoplasms / metabolism* COS Cells Carcinogenesis / genetics Cell Proliferation / genetics Chlorocebus aethiops Female Gene Expression Regulation, Neoplastic / drug effects Gene Expression Regulation, Neoplastic / genetics* Gene Knockout Techniques HEK293 Cells Humans Membrane Proteins / genetics Membrane Proteins / metabolism* Organoids / metabolism Phosphatidylinositol 3-Kinase / metabolism Protein Isoforms / genetics Protein Isoforms / metabolism Proto-Oncogene Proteins c-akt / metabolism Signal Transduction / drug effects Signal Transduction / genetics Smad Proteins / genetics Smad Proteins / metabolism* Transforming Growth Factor beta / pharmacology
IF 1.655
Human and Animal Cells 293(RCB1637) COS-7(RCB0539)