Effect Of LRIG3 Gene Overexpressing On Biological Character Of Glioma And The Mechanism

PartⅠEffect of LRIG3 gene overexpressing on proliferation of glioma U251MG and U87 cells and expression of PCNA and Ki67.Backgroud and Objective:This study is to explore the effect of over expression of LRIG3 gene on proliferation of glioma cells and expression of proliferating cell nuclear antigen(PCNA) and Ki67.Methods:The plasmid which containing LRIG3 gene and control were transduced into glioma U251MG and U87 cells respectively. The mRNA and protein levels of LRIG3 were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot. Cell proliferation was examined by MTT assay. The expressions of PCNA and Ki67 were detected by SABC.Result:Compared with control cell, the mRNA levels of LRIG3 was raised by 67.6% and 79.9% in LRIG3 cell of U87 and U251, respectively; Their protein levels were increased by 62.3%(U87) and 91.0%(U251) each. Over expression of LRIG3 resulted in the reduction of cell proliferation. The positive rate of PCNA was significantly lower in LRIG3 cells than in control cells[U87:33.6±4.82 VS 55.5±4.01, U251:27.49±3.17 VS 47.81±4.67 (p<0.05)]. The positive rate of Ki67 was also significantly decreased in LRIG3 transduced cells compared with control cells[U87 23.5±4.60VS 55.2±4.19,U251 24.3±3.76 VS 48.5±6.11 (p<0.01)].Conclusion:Up-regulating LRIG3 gene expression can reduce the proliferation of glioma U87 and U251 cells. PartⅡEffect of over expressed LRIG3 gene on cell cycle, adhesion, invasion and migration of glioma U251MG and U87 cells and the mechanical of signal pathwayObjective:This study is to explore the effect of over expression of LRIG3 gene on cell cycle, invasion and apoptosis of glioma U251MG and U87 cells, and to investigate possible mechanical of signal pathway.Methods:The plasmids LRIG3 and the blank plasmid as control were transduced into glioma U251MG and U87 cells. The mRNA and protein levels of LRIG3 were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot. Cell cycle was examined by flow cytometry (PI). Invasion was measured using Transwell assay and TDT-mediated dUTP-biotin nick end-labeling (TUNEL) were used to measure cell apoptosis.Result:Compared with control group, the mRNA level of LRIG3 was raised by 67.6% and 79.9% in LRIG3 cell of U251 and U87, respectively; Protein levels were increased by 134.1%(U251) and 3.3 folds (U87) each. Cell cycle analysis showed that over expressed LRIG3 cells were arrested in G0/G1 phase of U251MG (P<0.01). In Transwell assay, the average number of over expressed LRIG3 cells passing through the insert filter were decreased by 72.04%(U87) and 56.40%(U251) (p< 0.01).In TUNEL assay, over expressed LRIG3 cells were more apoptosis than the control cells.The expression level of c-myc was decreased, both p-ERK and p-Akt was decreased.Conclusion:Up-regulating LRIG3 gene expression can reduce the proliferation and differentiation, raise the cell apoptosis and decrease the invasion of glioma U87 and U251 cells via ERK and Akt pathway.