miRNAlipidomics.zip

We present here lipidomic data from HepG2 cell lysates treated with miRNA mimics (NT, miR-16, miR-30b, miR-30c). NT stands for non-targeting control siRNA, and miRNA mimics are ready-to-transfect mimics that behave like endogenous miRNAs. Lipid extraction was performed according to the method Bligh and Dyer. The following lipid species were added as internal standards: PC 14:0/14:0, PC 22:0/22:0, PE 14:0/14:0, PE 20:0/20:0 (di-phytanoyl), PS 14:0/14:0, PS 20:0/20:0 (di-phytanoyl), PI 17:0/17:0, LPC 13:0, LPC 19:0, LPE 13:0, Cer d18:1/14:0, Cer 17:0, D7-FC, CE 17:0, and CE 22:0. The residues were dissolved either in 10 mM ammonium acetate in methanol/chloroform (3:1, v/v) (for low mass resolution tandem mass spectrometry), or chloroform/methanol/2-propanol (1:2:4 v/v/v) with 7.5 mM ammonium formate (for high resolution mass spectrometry). The analysis of lipids was performed by direct flow injection analysis (FIA) using either a triple quadrupole mass spectrometer (FIA-MS/MS; QQQ triple quadrupole) or a hybrid quadrupole-Orbitrap mass spectrometer. Data processing was performed using the ALEX software which includes peak assignment and intensity picking. The extracted data were exported to Microsoft Excel 2010 and further processed by self-programmed Macros.本研究提交了经miRNA模拟物（miRNA mimics）处理的HepG2细胞裂解物的脂质组学数据，所涉处理组包括NT、miR-16、miR-30b及miR-30c；其中NT即非靶向对照小干扰RNA（non-targeting control siRNA），miRNA模拟物（miRNA mimics）为可直接转染、功能模拟内源性miRNA的预制制剂。脂质提取依照Bligh-Dyer法完成。本次实验添加了以下脂质物种作为内标物：磷脂酰胆碱（PC）14:0/14:0、磷脂酰胆碱（PC）22:0/22:0、磷脂酰乙醇胺（PE）14:0/14:0、磷脂酰乙醇胺（PE）20:0/20:0（二植烷酰基）、磷脂酰丝氨酸（PS）14:0/14:0、磷脂酰丝氨酸（PS）20:0/20:0（二植烷酰基）、磷脂酰肌醇（PI）17:0/17:0、溶血磷脂酰胆碱（LPC）13:0、溶血磷脂酰胆碱（LPC）19:0、溶血磷脂酰乙醇胺（LPE）13:0、神经酰胺（Cer）d18:1/14:0、神经酰胺（Cer）17:0、D7-胆固醇（D7-FC）、胆固醇酯（CE）17:0及胆固醇酯（CE）22:0。样品残渣采用两种溶剂体系复溶：其一为含10 mM乙酸铵的甲醇/三氯甲烷混合液（体积比3:1），用于低分辨串联质谱分析；其二为含7.5 mM甲酸铵的三氯甲烷/甲醇/2-丙醇混合液（体积比1:2:4），用于高分辨质谱分析。脂质分析采用直接流动注射分析（FIA）技术，分别搭载三重四极杆质谱仪（FIA-MS/MS；QQQ三重四极杆质谱仪）或混合型四极杆-轨道阱质谱仪完成。数据处理采用ALEX软件进行，该软件集成了峰识别与强度提取功能。提取得到的数据导出至Microsoft Excel 2010中，随后通过自行编写的宏完成后续处理。