Effect Of Liposomes And PEI Mediated PEGFP-N1-BmK CT Expression On The Migration Of Glioma Cells

BmK CT, isolated from the venom gland of Buthus martensii Karsch, is a 36 mer peptide cross-linked by four disulfide bridges. It has 68% sequence identity with CTX isolated from the scorpion Leiurus quinquestriatus. The biological character of BmK CT is a novel blocker of the chloride ion channel and MMP-2 of glioma cells.Matrix metalloproteinases (MMPs) are a family of endopeptidases that have the combined capacity to degrade all the components of the extracellular matrix. In this study, the mechanism of liposomes and PEI mediated a recombinant plasmid pEGFP-N1-BmK CT expression on the migration of glioma cell was analyzed at the cellular level. When recombinant plasmid pEGFP-N1-BmK CT was transfected into C6 glioma cells, the heterologous protein was expressed at a high level after 24 h. The secretion level of pro-MMP-2 and MMP-2 was suppressed in glioma cells harboring pEGFP-N1-BmK CT. Moreover, the migration rates of C6 glioma cells were inhibited in the pEGFP-Nl-BmK CT treatment group compared to that in the pEGFP-N1 treatment group. The MMP-2 was co-located with BmK CT around membrane of C6 cell nucleus.Lithium had been used clinically drugs in the treatment of bipolar disorder and was recently proved efficient in inhibiting migration and invasion of glioma cell. LiCl contributed to promote the inhibition of the secretion level of pro-MMP-2 and proliferation of pEGFP-N1-BmK CT-transfected glioma cells. When C6 cells were treated by LiCl, the accumulation of P-catenin in cell nucleus was increased in pEGFP-N1 group and pEGFP-N1-BmK CT group.Development of glioma-specific nanoparticles had been an area of intense research currently. Nanoparticles were investigated as drug-delivery vehicles and contrast agents. Surface modification of NPs with synthetic polymers such as polyethylenimine (PEI) would enhance the solubility of hydrophobic materials, biocompatibility, specific binding. PEI is a synthetic polymer that is commonly used as a gene vector because of its highly efficient transfection. In the present study, the effect of PEI mediated pEGFP-N1-BmK CT expression on the migration of glioma cell was analyzed. By using an Olympus FV1000 laser scanning confocal microscope, we observed that the expression of heterologous protein reached to 60% after 24 h transfection. The secretion level of pro-MMP-2 was tested in gelatin zymography assay. The activity of pro-MMP-2 was decreased in the BmK CT treatment group compared to that in the EGFP group. Furthermore, BmK CT displayed a higher activity in inhibiting cell migration than EGFP. Subsequently, four nanoparticles (ND-PEI, SiO2-PEI, MCM-41 and SBA-15) were synthesized. ND-PEI-DNA coupling rates increased gradually with the increased ratio of ND-PEI/DNA. In addition, size distribution of these NPs was determined by dynamic light scattering.The results reveal the antitumor efficacy of liposomes and PEI mediated BmK CT gene therapy, and provide a novel therapeutic strategy for glioma cancer.