Metabolomics Analysis in Non-Small Cell Lung Cancer by NMR Approach

Autor: Ming-Chi Yang, 楊明錡
Rok vydání: 2013
Druh dokumentu: 學位論文 ; thesis
Popis: 101
HLJ1, also known as DnaJB4, is a member of DnaJ-like heat shock protein 40 family. The previous studies had shown that HLJ1 expression is negatively correlated with invasion, migration, and proliferation in Non-Small Cell Lung Cancer (NSCLC). However, its role in metabolism is still unknown. Metabolomics has been vigorously developed in recent years, which is a powerful tool to investigate the changes in global metabolite profiles of cells. Therefore this study aims to set up the metabolomics approach and investigate the metabolite profiles of cancer cells with HLJ1 silence. First, the CL1-0 lung cancer cells were transduced with HLJ1 shRNA lentivirus to establish HLJ1-silenced stable cell clones. In addition, to get into the real situation of in vivo metabolomics, several organs from HLJ1 knockout mice, including lung, liver, kidney and heart, were applied to perform metabolomic analysis. Western blotting revealed that HLJ1 expression is significantly silenced no matter mice organs or cultured cell lysates. The metabolite profiles of each sample were detected using 1H nuclear magnetic resonance (NMR) spectrometry and analyzed by principal components analysis (PCA) for metabolite classification. In this study, we have successfully built up NMR-based analysis the metabolite profile of the organ and cell. We found that the classification of the hydrophilic material extracted by methanol is better than that of hydrophobic material by chloroform. Our results also showed that there are few differences in metabolomic profiles between HLJ1 expression and silence groups in mice, but no markedly difference in the cultured media and cell lysate extracts. Furthermore, the metabolomic results may combine with the microarray data of HLJ1-silenced cancer cells in the future, which might help us rapidly identify the targeted metabolic pathways that HLJ1 may be involved in.
Databáze: Networked Digital Library of Theses & Dissertations
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