Enhanced protection of Ins-1 β cells from apoptosis under hypoxia by delivery of DNA encoding secretion signal peptide-linked exendin-4 Exendin-4

Hyun Ah Kim, Suyeon Lee, Jeong Hyun Park, Sanghyun Lee, byungwan lee, Sung Hee Ihm, Tae Il Kim, Sung Wan Kim, Kyung Soo Ko, Minhyung Lee

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

In this study, we developed an expression system of exendin-4, a glucagon-like peptide (GLP-1) analog, using a secretion signal peptide (SP) to facilitate exendin-4 secretion. For delivery of the exendin-4 expression system, high-molecular-weight polyethylenimine (25 kDa, PEI25k), low-molecular-weight polyethylenimine (2 kDa, PEI2k), and polyamidoamine (PAMAM) dendrimers were evaluated as gene carriers to Ins-1 β cells. As a result, PEI25k showed the highest transfection efficiency. For the construction of the exendin-4 expression vector, DNA coding the SP sequence was inserted upstream of the exendin-4 cDNA, resulting in the construction of pβ-SP-Ex-4. Transfection assay showed that the secretion level of exendin-4 increased in the pβ-SP-Ex-4 transfected cells, compared with the pβ-Ex-4 transfected cells. To identify the β-cell protection effect of pβ-SP-Ex-4 delivery, the Ins-1 β cells were transfected with pβ-SP-Ex-4 or pβ-Ex-4 and incubated under normoxia or hypoxia. An MTT assay showed that the pβ-SP-Ex-4 transfected cells had higher β-cell viability than the pβ-Ex-4 transfected cells under hypoxia. In addition, the pβ-SP-Ex-4 transfected cells exhibited lower caspase-3 activity than the pβ-Ex-4 transfected cells. Therefore, PEI25k/pβ-SP-Ex-4 complex may be useful to protect isolated β cells from apoptosis during transplantation.

Original languageEnglish
Pages (from-to)242-248
Number of pages7
JournalJournal of Drug Targeting
Volume17
Issue number3
DOIs
Publication statusPublished - 2009 Apr 1

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Protein Sorting Signals
Apoptosis
DNA
Polyethyleneimine
Glucagon-Like Peptide 1
Transfection
Molecular Weight
exenatide
Hypoxia
Dendrimers
Cell Hypoxia
Cytoprotection
Caspase 3
Cell Survival
Complementary DNA
Transplantation

All Science Journal Classification (ASJC) codes

  • Pharmaceutical Science

Cite this

Ah Kim, Hyun ; Lee, Suyeon ; Park, Jeong Hyun ; Lee, Sanghyun ; lee, byungwan ; Hee Ihm, Sung ; Kim, Tae Il ; Wan Kim, Sung ; Soo Ko, Kyung ; Lee, Minhyung. / Enhanced protection of Ins-1 β cells from apoptosis under hypoxia by delivery of DNA encoding secretion signal peptide-linked exendin-4 Exendin-4. In: Journal of Drug Targeting. 2009 ; Vol. 17, No. 3. pp. 242-248.
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Enhanced protection of Ins-1 β cells from apoptosis under hypoxia by delivery of DNA encoding secretion signal peptide-linked exendin-4 Exendin-4. / Ah Kim, Hyun; Lee, Suyeon; Park, Jeong Hyun; Lee, Sanghyun; lee, byungwan; Hee Ihm, Sung; Kim, Tae Il; Wan Kim, Sung; Soo Ko, Kyung; Lee, Minhyung.

In: Journal of Drug Targeting, Vol. 17, No. 3, 01.04.2009, p. 242-248.

Research output: Contribution to journalArticle

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T1 - Enhanced protection of Ins-1 β cells from apoptosis under hypoxia by delivery of DNA encoding secretion signal peptide-linked exendin-4 Exendin-4

AU - Ah Kim, Hyun

AU - Lee, Suyeon

AU - Park, Jeong Hyun

AU - Lee, Sanghyun

AU - lee, byungwan

AU - Hee Ihm, Sung

AU - Kim, Tae Il

AU - Wan Kim, Sung

AU - Soo Ko, Kyung

AU - Lee, Minhyung

PY - 2009/4/1

Y1 - 2009/4/1

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AB - In this study, we developed an expression system of exendin-4, a glucagon-like peptide (GLP-1) analog, using a secretion signal peptide (SP) to facilitate exendin-4 secretion. For delivery of the exendin-4 expression system, high-molecular-weight polyethylenimine (25 kDa, PEI25k), low-molecular-weight polyethylenimine (2 kDa, PEI2k), and polyamidoamine (PAMAM) dendrimers were evaluated as gene carriers to Ins-1 β cells. As a result, PEI25k showed the highest transfection efficiency. For the construction of the exendin-4 expression vector, DNA coding the SP sequence was inserted upstream of the exendin-4 cDNA, resulting in the construction of pβ-SP-Ex-4. Transfection assay showed that the secretion level of exendin-4 increased in the pβ-SP-Ex-4 transfected cells, compared with the pβ-Ex-4 transfected cells. To identify the β-cell protection effect of pβ-SP-Ex-4 delivery, the Ins-1 β cells were transfected with pβ-SP-Ex-4 or pβ-Ex-4 and incubated under normoxia or hypoxia. An MTT assay showed that the pβ-SP-Ex-4 transfected cells had higher β-cell viability than the pβ-Ex-4 transfected cells under hypoxia. In addition, the pβ-SP-Ex-4 transfected cells exhibited lower caspase-3 activity than the pβ-Ex-4 transfected cells. Therefore, PEI25k/pβ-SP-Ex-4 complex may be useful to protect isolated β cells from apoptosis during transplantation.

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