Effects of Caspase-3 and of Glypicante1 altered expressions on proliferation of glioma cells and its mechanism

Abstract

Objective To explore the effects of Caspase-3 and Glypicante1 (Gpc1) altered expressions on the proliferation of glioma cells and its mechanism. Methods Fifty-two glioma specimens and 13 normal cerebral tissues were collected in our hospital from October 2011 to October 2014; the protein expressions of Caspase-3 and Gpc1 in these tissues were detected with immunohistochemistry. The peDNA3.1/Gpc1 high expression plasmids were constructed; human glioma A172 cell lines were routinely cultured in vitro; the signal path changes of Hedgehog (Hh) in the A172 cells after being transfected with 0.5, 1, 2 and 4 μg peDNA3.1/Gpc1 high expression plasmids were detected by Luciferase luciferase reporter gene kit. Gpc1-1694 interference plasmids were transfected into the A172 cells to silence the Gpc1 expression and these A172 cells were given 8 μg/mL Blasticidin (experimental group) for 24 h; normal A172 cells were given the same volume of solvent (control group); altered expressions of Gpc1 were detected by immumohistochemical staining and proliferation activity of the cells were observed by MTT assay 24, 48, 72 and 96 h after transfection. Results The expressions of Caspase-3 and Gpc1 in the glioma specimens were significantly higher than those in the normal brain tissues (P<0.05); in the glioma specimens, positive correlation between Caspase-3 and Gpc1 was noted (r=0.486, P=0.000). As compared with the control group, increased fluorescence intensity was observed in the 0.5, 1, 2 and 4 μg peDNA3.1/Gpc1 high expression plasmids groups (P< 0.05); immumohistochemical staining showed that CyclinD1 expression in the Hh pathway of in the peDNA 3.1/Gpc1 high expression plasmids groups was significantly lower than that in the control group; MTT assay showed that the proliferation activity of the A172 cells in the peDNA3.1/Gpc1 high expression plasmids groups at 24, 48, 72 and 96 h after transfection was significantly lower than that in the control group (P<0.05). Conclusion Caspase-3 and Gpc1 play important roles in regulating the proliferation of glioma cells, which might be related to Hedgehog signaling pathway. Key words: Caspase-3; Glypicante1; Glioma; Proliferation