Expression of Human Placenta-specific 1 (PLAC1) in CHO-K1 Cells
Copyright© 2020 Avicenna Research Institute..
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.
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:12 |
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| Enthalten in: |
Avicenna journal of medical biotechnology - 12(2020), 1 vom: 18. Jan., Seite 24-31 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Mahmoudian, Jafar [VerfasserIn] |
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| Themen: |
Eukaryotic cells |
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| Anmerkungen: |
Date Revised 13.04.2022 published: Print Citation Status PubMed-not-MEDLINE |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM307420388 |
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| 245 | 1 | 0 | |a Expression of Human Placenta-specific 1 (PLAC1) in CHO-K1 Cells |
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| 500 | |a Date Revised 13.04.2022 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status PubMed-not-MEDLINE | ||
| 520 | |a Copyright© 2020 Avicenna Research Institute. | ||
| 520 | |a 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 | ||
| 520 | |a 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) | ||
| 520 | |a 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 | ||
| 520 | |a CONCLUSION: These results indicated that there are some yet unknown PLAC1 localization signals employed by cancer cells for surface expression of PLAC1 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Eukaryotic cells | |
| 650 | 4 | |a PLAC1 protein expression | |
| 650 | 4 | |a Protein transport | |
| 700 | 1 | |a Nazari, Mahboobeh |e verfasserin |4 aut | |
| 700 | 1 | |a Ghods, Roya |e verfasserin |4 aut | |
| 700 | 1 | |a Jeddi-Tehrani, Mahmood |e verfasserin |4 aut | |
| 700 | 1 | |a Ostad, Seyed Nasser |e verfasserin |4 aut | |
| 700 | 1 | |a Ghahremani, Mohammad Hossein |e verfasserin |4 aut | |
| 700 | 1 | |a Vafaei, Sedigheh |e verfasserin |4 aut | |
| 700 | 1 | |a Amiri, Mohammad Mehdi |e verfasserin |4 aut | |
| 700 | 1 | |a Zarnani, Amir-Hassan |e verfasserin |4 aut | |
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