INHIBITION OF GLYCOLYSIS BY USING NANOLIPID BROMOPYRUVIC CHITOSAN CARRIER IS A PROMISING TOOL TO PREVENT HCC INVASIVENESS

Background and Aims: An important issue in nanotechnology is that the passage in solid tumour cells will be limited for carriers which have smaller diameter than the range of cancerous cells junctions (100-600nm). Thus, there is an urgent need to develop smart and efficient strategies tailoring size, shape, and specifically targeting to block one of glycolysis stage. In this way, folic acid (FA) as specific ligand for foliate receptor of cancer cells was conjugated with chitosan (CHI) which was attached with bovine serum albumin (BSA) as target of liver cells forming together (FA-CHI-BSA) to give rise to a specific carrier system. Our aim was to fabricate nano lipid carriers labelled by Rhodamine and covered by two layers: chitosan-bromopyruvic acid complex as one layer then conjugated by BSA-CHI-Folic acid as second layer.
Methods: Charge potential of lipid droplets and intensity of size distribution before and after sonication were investigated by photon correlation spectroscopy. Attachment of Bromopyruvic acid with chitosan was detected by infrared spectroscopy. Conjugation of folic acid and bovine serum albumin (BSA) with chitosan was also detected. Characterization of covering lipid droplets with chitosan-bromopyruvic acid was investigated by SEM, TEM and AFM before and after attachment. The effect of nano-lipid chitosan- BSA-FA encapsulated by bromopyruvic acid on the HCC growth was investigated upon time by crystal violet stain and MTT assay. Results: Bromopyruvic acid was attached electrostatically by chitosan through amino groups. Molecular fingerprint of bromopyruvic acid, chitosan and complex was detected. Nano- lipid droplets labeled by Rhodamine were covered by chitosan- bromopyruvic acid complex under rotation for 5 min. Afterwards folic BSA-CHI-FA complex was added by completing time of rotation up to 15min. Fluorescence microscopy TRITC and FITC channels revealed successful labelling of lipid droplets covered by complex. Morphological change of HCC growth was shown clearly by crystal violet stain. MTT assay indicates that bromopyruvic acid encapsulated has strongly affected HCC growth. Tryplan blue shows significant increase of apoptotic cells number upon time. Conclusions: Bromopyruvic acid has exhibited strong effect on HCC growth by blocking hexokinase II enzymes causing mitochondrial potential stress and reduction of ATP energy. Encapsulation of bromopyruvic acid in FA-BSA targeted nano-vehicle might offer protection of normal cell.