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 language | English (US) |
|---|---|
| Pages (from-to) | 1013-1021 |
| Number of pages | 9 |
| Journal | Acta Pharmacologica Sinica |
| Volume | 25 |
| Issue number | 8 |
| State | Published - Aug 1 2004 |
Fingerprint
Keywords
- Acetylcholinesterase
- Apoptosis
- Gene expression
- Proliferation
ASJC Scopus subject areas
- Pharmacology
- Pharmacology (medical)
Cite this
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 journal › Article
}
TY - JOUR
T1 - Overexpression of acetylcholinesterase inhibited cell proliferation and promoted apoptosis in NRK cells
AU - Jin, Qi Huang
AU - He, Heng Yi
AU - Shi, Yu Fang
AU - Lu, He
AU - Zhang, Xue Jun
PY - 2004/8/1
Y1 - 2004/8/1
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.
KW - Acetylcholinesterase
KW - Apoptosis
KW - Gene expression
KW - Proliferation
UR - http://www.scopus.com/inward/record.url?scp=4043104785&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=4043104785&partnerID=8YFLogxK
M3 - Article
C2 - 15301733
AN - SCOPUS:4043104785
VL - 25
SP - 1013
EP - 1021
JO - Zhongguo yao li xue bao = Acta pharmacologica Sinica
JF - Zhongguo yao li xue bao = Acta pharmacologica Sinica
SN - 1671-4083
IS - 8
ER -