Quantification of 5-methylcytosine and 5-hydroxymethylcytosine in genomic DNA from hepatocellular carcinoma tissues by capillary hydrophilic-interaction liquid chromatography/quadrupole TOF mass spectrometry.
Quantification of 5-methylcytosine and 5-hydroxymethylcytosine in genomic DNA from hepatocellular carcinoma tissues by capillary hydrophilic-interaction liquid chromatography/quadrupole TOF mass spectrometry.
复制标题
DOI：
10.1373/clinchem.2012.193938
复制
发表时间：
2013-05
影响因子：
9.3
通讯作者：
Yuan BF
中科院分区：
文献类型：
--
作者：
Chen ML;Shen F;Huang W;Qi JH;Wang Y;Feng YQ;Liu SM;Yuan BF
5-甲基胞嘧啶（5-mC）是参与发育的重要表观遗传修饰，并且经常在癌症中改变。5-mC可以酶促转化为5-羟甲基胞嘧啶（5-hmC）。5-已知hmC修饰在胚胎干细胞和神经元的DNA中是普遍的，但5-hmC在人肝肿瘤和匹配的对照组织中的分布尚未被严格探索。我们建立了一种在线捕获/毛细管电泳相互作用液相色谱（cHILIC）/源内裂解/串联质谱系统，用于定量测定肝细胞癌（HCC）肿瘤组织和相关癌旁组织基因组DNA中的5-mC和5-hmC。制备了亲水性聚合物整体柱，并将其用于cHILIC与在线捕集系统联用分离12种核苷。测定了该方法的检测限和定量限、回收率和不精密度。5-mC和5-hmC的检测限分别为0.06和0.19 fmol。测定了方法的不精密度和回收率，相对标准偏差<14.9%，相对误差<15.8%。HCC肿瘤组织的5-hmC含量比肿瘤相邻组织低4 - 5倍。此外，5-hmC含量与肿瘤分期高度相关（肿瘤淋巴结转移，P = 0.0002;巴塞罗那临床肝癌，P = 0.0003）。5-hmC的显著缺失可能对HCC的表观遗传调控产生深远影响，并可能成为HCC早期检测和预后的潜在生物标志物。
5-Methylcytosine (5-mC) is an important epigenetic modification involved in development and is frequently altered in cancer. 5-mC can be enzymatically converted to 5-hydroxymethylcytosine (5-hmC). 5-hmC modifications are known to be prevalent in DNA of embryonic stem cells and neurons, but the distribution of 5-hmC in human liver tumor and matched control tissues has not been rigorously explored. We developed an online trapping/capillary hydrophilic-interaction liquid chromatography (cHILIC)/in-source fragmentation/tandem mass spectrometry system for quantifying 5-mC and 5-hmC in genomic DNA from hepatocellular carcinoma (HCC) tumor tissues and relevant tumor adjacent tissues. A polymer-based hydrophilic monolithic column was prepared and used for the separation of 12 nucleosides by cHILIC coupled with an online trapping system. Limits of detection and quantification, recovery, and imprecision of the method were determined. Limits of detection for 5-mC and 5-hmC were 0.06 and 0.19 fmol, respectively. The imprecision and recovery of the method were determined, with the relative SDs and relative errors being <14.9% and 15.8%, respectively. HCC tumor tissues had a 4- to 5-fold lower 5-hmC content compared to tumor-adjacent tissues. In addition, 5-hmC content highly correlated with tumor stage (tumor-nodes-metastasis, P = 0.0002; Barcelona Clinic liver cancer, P = 0.0003). The marked depletion of 5-hmC may have profound effects on epigenetic regulation in HCC and could be a potential biomarker for the early detection and prognosis of HCC.
登录
查看更多内容
影响因子：
3.7
作者：
Globisch D;Münzel M;Müller M;Michalakis S;Wagner M;Koch S;Brückl T;Biel M;Carell T
通讯作者：
Carell T
一种敏感的质谱法，用于同时定量生物样品中的DNA甲基化和羟甲基水平。
影响因子：
2.9
作者：
Le T;Kim KP;Fan G;Faull KF
通讯作者：
Faull KF
影响因子：
11.2
作者：
Jin SG;Jiang Y;Qiu R;Rauch TA;Wang Y;Schackert G;Krex D;Lu Q;Pfeifer GP
通讯作者：
Pfeifer GP
DOI：
10.1126/science.1170116
发表时间：
2009-05-15
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Tahiliani M;Koh KP;Shen Y;Pastor WA;Bandukwala H;Brudno Y;Agarwal S;Iyer LM;Liu DR;Aravind L;Rao A
通讯作者：
Rao A
影响因子：
48
作者：
Song, Chun-Xiao;Clark, Tyson A.;Lu, Xing-Yu;Kislyuk, Andrey;Dai, Qing;Turner, Stephen W.;He, Chuan;Korlach, Jonas
通讯作者：
Korlach, Jonas