pEGFP-N1

The deleterious invasion of normal brain tissue by glioma cells is endorsed by its inherent ability to regulate the receptor-mediated adhesive properties, extracellular matrix degradation and remodeling, and elevated capacity for metalloproteinase (MMP) secretion (e.g., MMP-2). By doing so, it will create an intercellular space for glioma cells to invade. Here, researchers report that the gene therapy Buthus martensii Karsch (BmK) CT, a scorpion toxin peptide, combined with lithium chloride (LiCl), which is clinically used as a mood stabilizer, inhibits C6 glioma cell migration and Invasion. The results showed that simultaneous administration of LiCl and pEGFP-N1-BmK CT to glioma cells blocked the secretion of pro-MMP2 while inhibiting their proliferation in a synergistic manner.

In this study, the MTT method was used to detect the effect of LiCl on cell growth. Equal amounts of C6 glioma cells (10⁴ cells well^-1) were added to a 96-well microtiter plate, followed by medium to a final volume of 100 μl. Then, untreated cells were used as controls, or cells were transfected with pEGFP-N1 or pEGFP-N1-BmK CT and treated with LiCl for 24 hours. Tetrazolium salt was then added to each well, the cells were further incubated for 4 hours, the medium was discarded, and DMSO was added to each well. Record the absorbance of each well at 490 nm using a microplate reader. Generally speaking, LiCl is an inhibitor of GSK-3b and can inhibit cell proliferation activity. As shown in Figure 1, after pEGFPN1-BmK CT was transfected for 24 h and treated with LiCl for 24 h, LiCl treatment enhanced the growth inhibitory effect of pEGFP-N1-BmK CT. In both assays, the inhibition rate increased with pEGFP-N1-BmK CT and LiCl combined treatment compared with pEGFP-N1 and LiCl treatment.

Figure 1. Effect of lithium chloride and pEGFP-N1-BmK CT on C6 glioma cells proliferation. (Fu Y, et al., 2016)