Overexpression of acetylcholinesterase inhibited cell proliferation and promoted apoptosis in NRK cells

Qi Huang Jin, Heng Yi He, Yu Fang Shi, He Lu, Xue Jun Zhang

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

AIM: To study the potential function of acetylcholinesterase (AChE) in apoptosis through overexpression of AChE in Normal Rat Kidney (NRK) cells. METHODS: AChE activity was detected by the method of Karnovsky and Roots. Activated caspase-3 was analyzed by Western blotting and immunoflurescence with antibody special to activated caspase-3 fragment. The expression plasmids were constructed in pcDNA3.1 containing AChE gene or a fragment of AChE antisense that were got from RT-PCR. Stable expression cell lines were selected by G418 in cells transfected by lipofection. AChE expression was analyzed by RT-PCR and Western blotting. The proliferation rates of transfected cells were examined by the growth curve and cloning efficiency. MTT assay was used to analyze the cell viability. RESULTS: The proliferation rate of the cells transfected with AChE was retarded and the cloning efficiency was lower (28.2%±3.1% and 48.7%±2.1%) than cells transfected with vector (56.1%±0.3 %) or AChE-antisense (77.7%±2.2%). After 2 d the various clone types were deprived of serum, the residue cell viability were 10.4%±4.6% and 12.6%±6.7% in the cells transfected with AChE, and 27.4%±3.5% in cells with vector, and 50.3%±7.8% in cells with AChE-antisense. CONCLUSION: During apoptosis, increase of AChE protein is to inhibit cell proliferation, and then to promote apoptosis in NRK cells.

Original languageEnglish (US)
Pages (from-to)1013-1021
Number of pages9
JournalActa Pharmacologica Sinica
Volume25
Issue number8
StatePublished - Aug 1 2004

Fingerprint

Acetylcholinesterase
Cell Proliferation
Apoptosis
Kidney
Caspase 3
Organism Cloning
Cell Survival
Western Blotting
Polymerase Chain Reaction
Plasmids
Clone Cells
Cell Line
Antibodies

Keywords

  • Acetylcholinesterase
  • Apoptosis
  • Gene expression
  • Proliferation

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)

Cite this

Jin, Q. H., He, H. Y., Shi, Y. F., Lu, H., & Zhang, X. J. (2004). Overexpression of acetylcholinesterase inhibited cell proliferation and promoted apoptosis in NRK cells. Acta Pharmacologica Sinica, 25(8), 1013-1021.

Overexpression of acetylcholinesterase inhibited cell proliferation and promoted apoptosis in NRK cells. / Jin, Qi Huang; He, Heng Yi; Shi, Yu Fang; Lu, He; Zhang, Xue Jun.

In: Acta Pharmacologica Sinica, Vol. 25, No. 8, 01.08.2004, p. 1013-1021.

Research output: Contribution to journalArticle

Jin, QH, He, HY, Shi, YF, Lu, H & Zhang, XJ 2004, 'Overexpression of acetylcholinesterase inhibited cell proliferation and promoted apoptosis in NRK cells', Acta Pharmacologica Sinica, vol. 25, no. 8, pp. 1013-1021.
Jin, Qi Huang ; He, Heng Yi ; Shi, Yu Fang ; Lu, He ; Zhang, Xue Jun. / Overexpression of acetylcholinesterase inhibited cell proliferation and promoted apoptosis in NRK cells. In: Acta Pharmacologica Sinica. 2004 ; Vol. 25, No. 8. pp. 1013-1021.
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abstract = "AIM: To study the potential function of acetylcholinesterase (AChE) in apoptosis through overexpression of AChE in Normal Rat Kidney (NRK) cells. METHODS: AChE activity was detected by the method of Karnovsky and Roots. Activated caspase-3 was analyzed by Western blotting and immunoflurescence with antibody special to activated caspase-3 fragment. The expression plasmids were constructed in pcDNA3.1 containing AChE gene or a fragment of AChE antisense that were got from RT-PCR. Stable expression cell lines were selected by G418 in cells transfected by lipofection. AChE expression was analyzed by RT-PCR and Western blotting. The proliferation rates of transfected cells were examined by the growth curve and cloning efficiency. MTT assay was used to analyze the cell viability. RESULTS: The proliferation rate of the cells transfected with AChE was retarded and the cloning efficiency was lower (28.2{\%}±3.1{\%} and 48.7{\%}±2.1{\%}) than cells transfected with vector (56.1{\%}±0.3 {\%}) or AChE-antisense (77.7{\%}±2.2{\%}). After 2 d the various clone types were deprived of serum, the residue cell viability were 10.4{\%}±4.6{\%} and 12.6{\%}±6.7{\%} in the cells transfected with AChE, and 27.4{\%}±3.5{\%} in cells with vector, and 50.3{\%}±7.8{\%} in cells with AChE-antisense. CONCLUSION: During apoptosis, increase of AChE protein is to inhibit cell proliferation, and then to promote apoptosis in NRK cells.",
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AU - Jin, Qi Huang

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AU - Shi, Yu Fang

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N2 - AIM: To study the potential function of acetylcholinesterase (AChE) in apoptosis through overexpression of AChE in Normal Rat Kidney (NRK) cells. METHODS: AChE activity was detected by the method of Karnovsky and Roots. Activated caspase-3 was analyzed by Western blotting and immunoflurescence with antibody special to activated caspase-3 fragment. The expression plasmids were constructed in pcDNA3.1 containing AChE gene or a fragment of AChE antisense that were got from RT-PCR. Stable expression cell lines were selected by G418 in cells transfected by lipofection. AChE expression was analyzed by RT-PCR and Western blotting. The proliferation rates of transfected cells were examined by the growth curve and cloning efficiency. MTT assay was used to analyze the cell viability. RESULTS: The proliferation rate of the cells transfected with AChE was retarded and the cloning efficiency was lower (28.2%±3.1% and 48.7%±2.1%) than cells transfected with vector (56.1%±0.3 %) or AChE-antisense (77.7%±2.2%). After 2 d the various clone types were deprived of serum, the residue cell viability were 10.4%±4.6% and 12.6%±6.7% in the cells transfected with AChE, and 27.4%±3.5% in cells with vector, and 50.3%±7.8% in cells with AChE-antisense. CONCLUSION: During apoptosis, increase of AChE protein is to inhibit cell proliferation, and then to promote apoptosis in NRK cells.

AB - AIM: To study the potential function of acetylcholinesterase (AChE) in apoptosis through overexpression of AChE in Normal Rat Kidney (NRK) cells. METHODS: AChE activity was detected by the method of Karnovsky and Roots. Activated caspase-3 was analyzed by Western blotting and immunoflurescence with antibody special to activated caspase-3 fragment. The expression plasmids were constructed in pcDNA3.1 containing AChE gene or a fragment of AChE antisense that were got from RT-PCR. Stable expression cell lines were selected by G418 in cells transfected by lipofection. AChE expression was analyzed by RT-PCR and Western blotting. The proliferation rates of transfected cells were examined by the growth curve and cloning efficiency. MTT assay was used to analyze the cell viability. RESULTS: The proliferation rate of the cells transfected with AChE was retarded and the cloning efficiency was lower (28.2%±3.1% and 48.7%±2.1%) than cells transfected with vector (56.1%±0.3 %) or AChE-antisense (77.7%±2.2%). After 2 d the various clone types were deprived of serum, the residue cell viability were 10.4%±4.6% and 12.6%±6.7% in the cells transfected with AChE, and 27.4%±3.5% in cells with vector, and 50.3%±7.8% in cells with AChE-antisense. CONCLUSION: During apoptosis, increase of AChE protein is to inhibit cell proliferation, and then to promote apoptosis in NRK cells.

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