【摘 要】
：
5-methylcytosine(5-mC)has long been known to be present in RNA from all three kingdoms of life.However,the functions of 5-mC in RNA havent been fully unders
【机 构】
：
KeyLaboratoryofAnalyticalChemistryforBiologyandMedicine(MinistryofEducation),DepartmentofChemistry,W
【出 处】
：
2016年分析化学前沿国际研讨会及中美分析化学研讨会
论文部分内容阅读
5-methylcytosine(5-mC)has long been known to be present in RNA from all three kingdoms of life.However,the functions of 5-mC in RNA havent been fully understood,especially for RNA demethylation mechanism.The discovery of 5-hydroxymethylcytosine(5-hmC)in RNA together with our recently reported 5-formylcytosine(5-foC)in RNA [1] indicated 5-mC in RNA may undergo the same cytosine oxidation demethylation pathway with generating the intermediates of 5-hmC,5-foC,and 5-carboxylcytosine(5-caC)by ten-eleven translocation(Tet)proteins.However,endogenous 5-caC in RNA hasnt been observed so far.Here we established a method by chemical labeling coupled with liquid chromatography-mass spectrometry analysis for sensitive and simultaneous determination of the oxidative products of 5-mC.Our results demonstrated that the detection sensitivities of 5-mC,5-hmC,5-foC and 5-caC in RNA increased by 70-313 folds upon 2-bromo-1-(4-diethylaminophenyl)-ethanone(BDEPE)labeling.Using this method,we discovered the existence of 5-caC in RNA of mammals [2].In addition,we found 5-mC occurs in all RNA species including mRNA,28S rRNA,18S rRNA and small RNA(< 200 nt).However,5-hmC,5-foC and 5-caC mainly occur in mRNA,and barely detected in other types of RNA.We also found that the contents of 5-hmC in RNA of human colorectal carcinoma(CRC)and hepatocellular carcinoma(HCC)tissues significantly decreased compared to tumor adjacent normal tissues,suggesting that 5-hmC in RNA may play certain functional roles in the regulation of cancer development and formation.
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