A Novel Antisense Oligodeoxynucleotide Targeting Caspase-3 mRNA Prevents PC-12 Cell Apoptosis Induced by Aβ_(25-35)
Ya Wang
Abstract
Ya Wang
Abstract
Objective To design and synthesize antisense oligodeoxynucleotides (ASODNs) targeting caspase 3 mRNA and to screen the effective ASODNs to prevent PC 12 cells apoptosis induced by Aβ 25 35 Methods 17 18 mer ASODNs targeting different regions of caspase 3 mRNA were designed and transferred into PC 12 cells with Lipofectimine and followed by induction with Aβ 25 35 The effective ASODN preventing PC 12 cells from apoptosis was screened according to the results of Flow Cytometry (FCM) analysis with annexinⅤ FITC/Propidium Iodide (PI) dual staining. Results FCM analysis by annexinⅤ/PI staining showed that the ratio of apoptotic cells decreased more significantly than other ODNs after the ASODN 2 ( GTT GTT GTC CAT GGT CAC )(P0 05), which targets bases 6 12 of caspase 3 mRNA and was transferred into PC 12 cells , then the cells were exposed to Aβ 25 35 Apoptotic cells transferred with ASODN\-2 treated with Aβ 25 35 were different from Aβ 25 35 group without statistical significance. Conclusion Caspase 3 plays an important role in the PC 12 cells apoptosis treated with Aβ 25 35 The ASODN 2 aimed at caspase 3 mRNA was identified as an effective fragment to prevent the PC 12 cells from apoptosis treated with Aβ 25 35 .
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Objective To design and synthesize antisense oligodeoxynucleotides (ASODNs) targeting caspase 3 mRNA and to screen the effective ASODNs to prevent PC 12 cells apoptosis induced by Aβ 25 35 Methods 17 18 mer ASODNs targeting different regions of caspase 3 mRNA were designed and transferred into PC 12 cells with Lipofectimine and followed by induction with Aβ 25 35 The effective ASODN preventing PC 12 cells from apoptosis was screened according to the results of Flow Cytometry (FCM) analysis with annexinⅤ FITC/Propidium Iodide (PI) dual staining. Results FCM analysis by annexinⅤ/PI staining showed that the ratio of apoptotic cells decreased more significantly than other ODNs after the ASODN 2 ( GTT GTT GTC CAT GGT CAC )(P0 05), which targets bases 6 12 of caspase 3 mRNA and was transferred into PC 12 cells , then the cells were exposed to Aβ 25 35 Apoptotic cells transferred with ASODN\-2 treated with Aβ 25 35 were different from Aβ 25 35 group without statistical significance. Conclusion Caspase 3 plays an important role in the PC 12 cells apoptosis treated with Aβ 25 35 The ASODN 2 aimed at caspase 3 mRNA was identified as an effective fragment to prevent the PC 12 cells from apoptosis treated with Aβ 25 35 .
Key concepts: Apoptosis, Propidium iodide, Annexin, Molecular biology, Flow cytometry, Messenger RNA, Pi, Biology