BioProject

In most solid tumors, the Warburg effect, also known as aerobic glycolysis, represents a major biochemical alteration associated with malignant transformation. More...

In most solid tumors, the Warburg effect, also known as aerobic glycolysis, represents a major biochemical alteration associated with malignant transformation. Although the exact molecular mechanisms underlying this metabolic change remain to be clarified, the biochemical alteration in cancer cell energy metabolism opens novel avenues for the development of therapeutic strategies to preferentially kill cancer cells by targeting the glycolytic pathway. We used microarrays to focus on differences among A375, M6 melanoma cells and normal melanocytes (HeMa cells) in terms of glucose metabolism- and invasion-related properties. The transcript analysis showed that melanoma cells exhibit typical transcriptional features of a glycolytic phenotype, matched with high invasive markers (including increased expression of several matrix metallo-proteinases) Overall design: Primary Human Epidermal Melanocytes (HeMa) isolated from adult skin were purchased from GIBCO Life Technologies Corporation (California, USA) and were grown in Medium 254CF with calcium plus Human Melanocytes Growth Supplement, obtained from the same company, were used as control . The melanoma cell line A375 (MITF wild type, BRAF V600E, NRAS wild type) was obtained from American Type Culture Collection and grown in Dulbecco’s modified Eagle’s medium containing 2 mM glutamine, 100 UI/ml penicillin, 100 μg/ml streptomycin and supplemented with 10 % FBS (Euroclone, Milano, Italy). cDNA samples have been generated and labeled from three independent RNA samples of each cellular line. Principal Component Analysis confirmed no batch effect among samples, i.e. no technical sources of variation that have been added to the samples during handling A375-M6 melanoma cells (M6) were isolated in our laboratory from lung metastasis of SCID bg/bg mice i.v. injected with A375 human melanoma cell line. A375 cell line was independently validated by STR profiling by the DNA diagnostic centre BMR Genomics (Padova, Italy). The cellular samples for array studies were amplified and kept in culture as mentioned above. Three chips for each experimental group (normal melanocytes, A375 cells and A375-M6 cells) were used in the microarray experiments. Less...