ABSTRACT. Isolation of high-quality RNA free of contaminants, such as polyphenols, proteins, plant secondary metabolites, and genomic DNA from plant tissues, is usually a challenging but crucial step for molecular analysis. We developed a novel protocol based on the cetyltrimethylammonium bromide method to isolate high-quality RNA from blackberry plant tissues, especially fruits. Most DNA was removed when acetic acid was utilized, before RNA precipitation. Thus, lithium chloride, a reagent widely used for RNA purification, was not needed. The isolation time was shortened to less than 3 h. The RNA was quite pure, with little DNA contamination. The quality of the RNA was assessed by spectrophotometric readings and electrophoresis on agarose gels. It was good enough for downstream enzymatic reactions, such as reverse transcription-PCR, cloning and real-time PCR assay. The method yielded an amount of total RNA comparable to previously described protocols.

Tumor cells undergo metabolic reprogramming to meet their energy and anabolic demands to maintain their malignant phenotype. Activation of oncogenes and deletion of tumor suppressors promotes metabolic reprogramming in cancer by directly or indirectly regulating enzymatic activities associated with metabolic pathways. Metabolic reprogramming in tumor cells mainly involves the glycolytic pathway, pentose phosphate pathway, serine synthesis pathway, enhanced glutamine metabolism or fatty acid anabolism, and abnormal mitochondrial oxidative phosphorylation (OXPHOS). The tricarboxylic acid (TCA) cycle is the central pathway of mitochondrial OXPHOS, and glucose, amino acid and fatty acid metabolism are associated with the TCA cycle. Metabolic abnormalities and rewiring of metabolic pathways are also present in Osteosarcoma (OS). The abnormal metabolic pattern in OS is associated with cell proliferation, migration, invasion and drug resistance. This review summarizes the current studies on glycolysis, amino acid metabolism, lipid synthesis and the TCA cycle related to OS.

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