1H-NMR-based metabolomics of tumor tissue for the metabolic characterization of rat hepatocellular carcinoma formation and metastasis

Abstract

The high mortality figures for hepatocellular carcinoma (HCC) are mostly due to high recurrence and metastasis rates. However, the metabolic characteristics of HCC metastasis have not been studied extensively. In this study, we attempted to elucidate the metabolite profile of HCC formation and metastasis through proton nuclear magnetic resonance ((1)H-NMR)-based metabolomics. We first established a hepatocellular carcinoma with lung metastasis (HLM) rat model by exposure to diethylnitrosamine. Fifteen rats were then divided into three groups based on pathologic changes: HCC, HLM, and controls. The metabolite profiles of extracts from tumor tissue were obtained using high-resolution (1)H-NMR. One-way ANOVA was used to compare the metabolite levels among the three groups. Multivariate statistical analysis (specifically, unsupervised principal components analysis and supervised partial least squares discriminant analysis (PLS-DA)) were used for HCC and HLM metabolite profiling and data interpretation. PLS-DA models could discern HCC or HLM rats from normal (control) rats. Tumor tissue from HLM showed changes in glucose, lactate, choline, lipids, and some amino acids such as glycine. The results of this pilot study suggest that alterations in glycolysis and the metabolism of glycine and choline occur during HCC invasion and metastasis.

The metabolomic window into hepatobiliary disease

“…Many researchers, particularly in China, have employed metabolomic protocols towards this end. Table 4 contains details of 24 metabolomic investigations of human HCC [27, 32, 36–57], three of chemically-induced rat HCC [42, 46, 58] and two of hepatocellular adenomas in the flatfish Limanda limanda [59, 60]. Many investigators of human HCC employed healthy volunteers as a control group, especially for the collection of serum/plasma or urine [27, 32, 36, 37, 39–41, 43–45, 47–50, 53, 57], others used cirrhotics as a comparator group [36, 37, 39, 45–48, 50–52, 54], while others included acute hepatitis [36, 37], chronic hepatitis [36, 37, 46, 48, 50], benign liver tumours [43] and acute myeloid leukemia [45] as comparator groups.…”

“…These metabolomic comparisons have permitted insights into the biochemical transitions to HCC from various precursor states, at least as viewed through serum/plasma or urine. A relatively few studies have addressed the hepatic metabolome directly by interrogating tumour tissue and paired uninvolved liver, for human HCC [38, 55, 56], chemically-induced rat HCC [42] and fish hepatocellular adenoma [59, 60]. Two recent reports also combined transcriptomic and metabolomic analyses of human HCC [55, 56].…”

Turmeric enhancing anti-tumor effect of Rhizoma paridis saponins by influencing their metabolic profiling in tumors of H22 hepatocarcinoma mice

“…A c c e p t e d M a n u s c r i p t 4 reproducible, and cheap methods acquires metabolic profiles in biological specimens without extensive sample preparation [2,30].…”

“…The aqueous extract sample spectra contained signals of carbohydrate, amino acids and organic acids, which were identified and labeled in the spectra, mainly according to Chenomx Profiler 7.6 identification and data from the literature [7,23,29,30]. These spectra data are given in detail in Table 2 …”

Quantification of Water-Soluble Metabolites in Medicinal Mushrooms Using Proton NMR Spectroscopy

“…[13][14][15] This complicated task can be effectively accomplished by combining NMR spectroscopy with metabolite identification and quantification in NMR spectra using a technique called "targeted profiling. "…”