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Down-regulation amyloid beta-protein 42 production by interfering with transcript of presenilin 1 gene with siRNA.
Acta Pharmacol Sin. 2004 Dec; 25(12):1613-8.AP

Abstract

AIM

To investigate the pathogenesis of Abeta42 yielding and new drug targets as well as the possibility of RNA interference (RNAi) technique for treatment of Alzheimer disease (AD).

METHODS

Human AD presenilin 1 (PS1) cDNA sequence was obtained from NCBI website. The three sites of RNAi action and one missense control site were selected in PS1 cDNA through online design of Ambion company. To confirm specificity of these sites, we conducted a BLAST search of the IMAGE EST library. The corresponding double-stranded DNA was used to construct pSilencer 3.1-H1 plasmid, which could transcribe small interference RNA (siRNA). Then, the pSliencer 3.1-H1 plasmids were transfected into CHO/PS1/APP cells with SuperFect transfection reagent. The cells have been transfected with the mutant PS1 and APP gene of AD. All the CHO/PS1/APP cells with pSliencer 3.1-H1 plasmids were screened out using G418. Transcripts of PS1 gene in CHO/PS1/APP were measured by RT-PCR, the contents of PS1 peptide and Abeta42 production inside CHO/PS1/APP cells were examined through Western blot and the Abeta42 change of secretion by CHO/PS1/APP was determined with ELISA.

RESULTS

The site 3 of PS1 mRNA was inhibited by RNAi after 2 d. The effect was more obvious with the time. The peptide corresponding to PS1 gene and Abeta42 production in CHO/PS1/APP cells were both reduced after siRNA interfere for 3 d. Abeta42 secretion by CHO/PS1/APP cells began to reduce on d 3, and reached the most significance on d 5. There was a time-dependent relationship between the transcript of PS1 gene and the production of Abeta42 with RNAi action.

CONCLUSION

PS1 is essential for g-secretase activity. Inhibition of the PS1 can decrease the levels of Abeta42. Some sites of PS1 mRNA, for example, the site 3, may serve as a new drug target and RNAi probably can be used for treatment of AD.

Authors+Show Affiliations

Neuropharmacological Research Laboratory, College of Pharmacy, Ji-nan University, Guangzhou 510632, China. tlhm@jnu.edu.cnNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15569405

Citation

Luo, Huan-min, et al. "Down-regulation Amyloid Beta-protein 42 Production By Interfering With Transcript of Presenilin 1 Gene With SiRNA." Acta Pharmacologica Sinica, vol. 25, no. 12, 2004, pp. 1613-8.
Luo HM, Deng H, Xiao F, et al. Down-regulation amyloid beta-protein 42 production by interfering with transcript of presenilin 1 gene with siRNA. Acta Pharmacol Sin. 2004;25(12):1613-8.
Luo, H. M., Deng, H., Xiao, F., Gao, Q., Weng, W., Zhang, P. F., & Li, X. G. (2004). Down-regulation amyloid beta-protein 42 production by interfering with transcript of presenilin 1 gene with siRNA. Acta Pharmacologica Sinica, 25(12), 1613-8.
Luo HM, et al. Down-regulation Amyloid Beta-protein 42 Production By Interfering With Transcript of Presenilin 1 Gene With SiRNA. Acta Pharmacol Sin. 2004;25(12):1613-8. PubMed PMID: 15569405.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Down-regulation amyloid beta-protein 42 production by interfering with transcript of presenilin 1 gene with siRNA. AU - Luo,Huan-min, AU - Deng,Hui, AU - Xiao,Fei, AU - Gao,Qin, AU - Weng,Wen, AU - Zhang,Pei-fen, AU - Li,Xiao-guang, PY - 2004/12/1/pubmed PY - 2005/6/25/medline PY - 2004/12/1/entrez SP - 1613 EP - 8 JF - Acta pharmacologica Sinica JO - Acta Pharmacol Sin VL - 25 IS - 12 N2 - AIM: To investigate the pathogenesis of Abeta42 yielding and new drug targets as well as the possibility of RNA interference (RNAi) technique for treatment of Alzheimer disease (AD). METHODS: Human AD presenilin 1 (PS1) cDNA sequence was obtained from NCBI website. The three sites of RNAi action and one missense control site were selected in PS1 cDNA through online design of Ambion company. To confirm specificity of these sites, we conducted a BLAST search of the IMAGE EST library. The corresponding double-stranded DNA was used to construct pSilencer 3.1-H1 plasmid, which could transcribe small interference RNA (siRNA). Then, the pSliencer 3.1-H1 plasmids were transfected into CHO/PS1/APP cells with SuperFect transfection reagent. The cells have been transfected with the mutant PS1 and APP gene of AD. All the CHO/PS1/APP cells with pSliencer 3.1-H1 plasmids were screened out using G418. Transcripts of PS1 gene in CHO/PS1/APP were measured by RT-PCR, the contents of PS1 peptide and Abeta42 production inside CHO/PS1/APP cells were examined through Western blot and the Abeta42 change of secretion by CHO/PS1/APP was determined with ELISA. RESULTS: The site 3 of PS1 mRNA was inhibited by RNAi after 2 d. The effect was more obvious with the time. The peptide corresponding to PS1 gene and Abeta42 production in CHO/PS1/APP cells were both reduced after siRNA interfere for 3 d. Abeta42 secretion by CHO/PS1/APP cells began to reduce on d 3, and reached the most significance on d 5. There was a time-dependent relationship between the transcript of PS1 gene and the production of Abeta42 with RNAi action. CONCLUSION: PS1 is essential for g-secretase activity. Inhibition of the PS1 can decrease the levels of Abeta42. Some sites of PS1 mRNA, for example, the site 3, may serve as a new drug target and RNAi probably can be used for treatment of AD. SN - 1671-4083 UR - https://www.unboundmedicine.com/medline/citation/15569405/Down_regulation_amyloid_beta_protein_42_production_by_interfering_with_transcript_of_presenilin_1_gene_with_siRNA_ L2 - http://www.chinaphar.com/1671-4083/25/1613.pdf DB - PRIME DP - Unbound Medicine ER -