An increasing data indicates that altered microRNAs(miRNAs)participate in the radiation-induced DNA damage response.However,a correlation of mRNA and miRNA profiles across the entire genome and in response to irradiat...An increasing data indicates that altered microRNAs(miRNAs)participate in the radiation-induced DNA damage response.However,a correlation of mRNA and miRNA profiles across the entire genome and in response to irradiation has not been thoroughly assessed.We analyzed miRNA microarray data collected from HeLa cells after ionizing radiation(IR),quantified the expression profiles of mRNAs and performed comparative analysis of the data sets using target prediction algorithms,Gene Ontology(GO)analysis,pathway analysis,and gene network construction.The results showed that the altered miRNAs were involved in regulation of various cellular functions.miRNA-gene network analyses revealed that miR-186,miR-106b,miR-15a/b,CCND1and CDK6 played vital role in the cellular radiation response.Using qRT-PCR,we confirmed that twenty-two miRNAs showed differential expression in HeLa cells treated with IR and some of these miRNAs affected cell cycle progression.This study demonstrated that miRNAs influence gene expression in the entire genome during the cellular radiation response and suggested vital pathways for further research.展开更多
Ionizing radiation (IR) causes severe cellular damage both directly and indirectly and disrupts RNA integrity. RNA strand breaks are the most frequent type of damage caused by IR. RNA damage is involved in the develop...Ionizing radiation (IR) causes severe cellular damage both directly and indirectly and disrupts RNA integrity. RNA strand breaks are the most frequent type of damage caused by IR. RNA damage is involved in the development of degenerative diseases, including Alzheimer’s disease and Parkinson’s disease. However, the mechanism of mRNA damage and any resulting pathophysiological outcomes are poorly understood. This is partly because there is a lack of sensitive tools to monitor damage randomly occurring in RNA, especially RNA strand break damage in a given RNA. In this work, a method using the reverse transcription polymerase chain reaction (RT-PCR) after poly(A) addition to 3′-end of RNA to determine RNA strand break damage in a specific RNA by poly(A) polymerase has been developed. The levels of damage in specific mRNAs, including ABL1, TP53, GADD45A and ATR from IR-treated HeLa cells were examined. Strand breaks were detected in all mRNAs examined. The study provides a novel and sensitive method based on 3 -end poly(A)-tailing RT-PCR to monitor RNA strand break damage.展开更多
基金supported by the National Basic Research Program of China(2010CB912801 and 2007CB914601)the National Natural Science Foundation of China(31170713)the Beijing Municipal Natural Science Foundation(5112029)
文摘An increasing data indicates that altered microRNAs(miRNAs)participate in the radiation-induced DNA damage response.However,a correlation of mRNA and miRNA profiles across the entire genome and in response to irradiation has not been thoroughly assessed.We analyzed miRNA microarray data collected from HeLa cells after ionizing radiation(IR),quantified the expression profiles of mRNAs and performed comparative analysis of the data sets using target prediction algorithms,Gene Ontology(GO)analysis,pathway analysis,and gene network construction.The results showed that the altered miRNAs were involved in regulation of various cellular functions.miRNA-gene network analyses revealed that miR-186,miR-106b,miR-15a/b,CCND1and CDK6 played vital role in the cellular radiation response.Using qRT-PCR,we confirmed that twenty-two miRNAs showed differential expression in HeLa cells treated with IR and some of these miRNAs affected cell cycle progression.This study demonstrated that miRNAs influence gene expression in the entire genome during the cellular radiation response and suggested vital pathways for further research.
基金supported by the National Basic Research Program of China (2007CB914601, 2010CB912801)Beijing Municipal Natural Science Foundation (5112029)
文摘Ionizing radiation (IR) causes severe cellular damage both directly and indirectly and disrupts RNA integrity. RNA strand breaks are the most frequent type of damage caused by IR. RNA damage is involved in the development of degenerative diseases, including Alzheimer’s disease and Parkinson’s disease. However, the mechanism of mRNA damage and any resulting pathophysiological outcomes are poorly understood. This is partly because there is a lack of sensitive tools to monitor damage randomly occurring in RNA, especially RNA strand break damage in a given RNA. In this work, a method using the reverse transcription polymerase chain reaction (RT-PCR) after poly(A) addition to 3′-end of RNA to determine RNA strand break damage in a specific RNA by poly(A) polymerase has been developed. The levels of damage in specific mRNAs, including ABL1, TP53, GADD45A and ATR from IR-treated HeLa cells were examined. Strand breaks were detected in all mRNAs examined. The study provides a novel and sensitive method based on 3 -end poly(A)-tailing RT-PCR to monitor RNA strand break damage.
