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Expression of Human Placenta-Specific 1 (Plac1) in Cho-K1 Cells



Mahmoudian J1, 2 ; Nazari M2 ; Ghods R3, 4 ; Jedditehrani M2 ; Ostad SN1, 5 ; Ghahremani MH1, 5 ; Vafaei S6 ; Amiri MM7 ; Zarnani AH6, 7, 8
Authors
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Authors Affiliations
  1. 1. Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences (TUMS), Tehran, Iran
  2. 2. Monoclonal Antibody Research Center, Avicenna Research Institute (ACECR), Tehran, Iran
  3. 3. Oncopathology Research Center, Iran University of Medical Sciences (IUMS), Tehran, Iran
  4. 4. Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences (IUMS, Tehran, Iran
  5. 5. Department of Toxicology and Pharmacology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
  6. 6. Reproductive Immunology Research Center, Avicenna Research Institute, (ACECR), Tehran, Iran
  7. 7. Department of Immunology, School of Public Health, Tehran University of Medical Sciences (TUMS), Tehran, Iran
  8. 8. Immunology Research Center (IRC), Iran University of Medical Sciences (IUMS, Tehran, Iran

Source: Avicenna Journal of Medical Biotechnology Published:2020

Abstract

Background: Placenta-specific 1 (PLAC1), as a new Cancer/Testis Antigen (CTA), is frequently expressed in a variety of cancers and localized to cytoplasm and plasma membrane. Surface expression of cancer target antigens is of great importance that enables antibody-mediated cancer immunotherapy. The aim of the current study was to express the intact human PLAC1 protein on plasma membrane of a eukaryotic cell as a model for future anti-PLAC1-based cancer immunotherapy. Methods: In the first approach, entire human PLAC1 gene including its own Signal Peptide (SP) was cloned into pIRES2-EGFP and LeGO-iG2 vectors and expressed in CHO-K1 cells. In the second approach, cytosolic and Signal-Anchor (SA) sequence of Transferrin Receptor Protein 1 (TFR1) were fused to extracellular portion of PLAC1 and expressed as above. Expression of PLAC1 was then assessed using Reverse Transcription Polymerase Chain Reaction (RT-PCR), Western Blot (WB), Immunocytochemistry (ICC), Immunofluorescence (IF) and Flow Cytometry (FC). Results: The first approach resulted in the expression of PLAC1 in submembranous but not in the surface of transfected CHO-K1 cells. Using the chimeric human PLAC1 construct, the same intracellular expression pattern was observed. Conclusion: These results indicated that there are some yet unknown PLAC1 localization signals employed by cancer cells for surface expression of PLAC1. © 2020, Avicenna Journal of Medical Biotechnology. All rights reserved.
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