The Experimental Study Of The Molecular Biological Mechanism Of Leucine-rich Repeats And Immunoglobulin-like Domains 3 (LRIG3) On The Biological Characterization Of Glioblastoma Cell Line GL15

Partâ… The Experimental Study of Relevant Biological Characterization of Glioblastoma Cell Line GL15Objective To explicate the biological character of glioblastoma cell line GL15 and to study the effect of EGF to the biological character of GL15 cell line.Methods The morphology, growth and proliferation of the glioblastoma cell line GL15 was observed during continuous culture in vitro. The karyotype of GL15 cells with Gimsa stain was analyzed. The expression of glial fibrillary acidic protein(GFAP) and proliferating cell nuclear antigen(PCNA) protein were detected with immuocytochemistry, the cell cycle was detected by flow cytometry(FCM), the effect of EGF on the biological character of GL15 cell line was measured by methyl thiazolyl tetrazolium (MTT) and Transwell, and campared with these characterizations of U251MG and U373MG glioblastoma cell line.Results N0. 7 chromosome, EGFR located, is over amplification with Gimsa stain. The morphology of GL15 cells are typical glioblastoma cell morphology. They grew and proliferated well in cell culture condition and were able to invade and form cell clone in vitro. The expression of GFAP and PCNA are positive, mainly in kytoplasm and nuclei respectively, and we can find many mitotic nucleus cells under microscopy. EGF can regulate the cell proliferation, invasive ability and GFAP expression of GL15.Conclusion GL15 cell line remains the biological character of typical glioblastoma cell character, it is an ideal cell platform to study glioblastoma character. Its biological characterization are intimate correlated to EGFR.Partâ…¡EGF and AG1478 affect the proliferation, invasiveness and GFAP expression of GL15 cell lineObjective To investigate the molecular mechanisms of epidermal growth factor receptor(EGFR) signal pathway affecting the biology characters of glioblastoma cell lines GL15.Methods The glioblastoma cell lines GL15 was cultured. The GL15 proliferation and invasive ability were detected by methyl thiazolyl tetrazolium (MTT) and Transwell, the expression of glial fibrillary acidic protein(GFAP) mRNA and protein were detected with reverse transcriptase-polymerase chain reaction(RT-PCR) and western blot after epidermal growth factor(EGF) and AG1478 intervention respectively.Results Ectogenic EGF can regulate the proliferation and invasiveness, small dose (50ng/ml,100ng/ml) promote the proliferation and enhance the invasiveness, while high dose (200ng/ml) step dowm the proliferation and invasiveness, AG1478 can inhibit the biological activity of EGFR.Conclusion The biological characteristics of glioblastoma cell lines GL15 are intimate correlated to EGFR. EGF regulate the growth of GL15 in two-way and AG1478 can inhibit the biological activity of EGFR non-completely. EGFR may be the ideal therapeutic target with gene methods. Partâ…¢The Molecular Mechanism Study of LRIG3 Regulated Cytobiology Activity of GL15 Cell LineObjective To investigate the molecular mechanisms of Leucine-rich Repeats and Immunoglobulin-like Domains 3(LRIG3)on the biological activity of glioblastoma cell lines GL15.Methods The glioblastoma cell lines GL15 was cultured and transfected with LRIG3-EGFP plasmid, then analyzed the change of the cell cycle after transfected, The proliferation and invasiveness were detected after transfected with methyl thiazolyl tetrazolium (MTT) and Transwell, the expression of EGFR and LRIG3 mRNA and protein were detected with reverse transcriptase-polymerase chain reaction(RT-PCR) and western blot after transfected respectively.Results After transfected with the plasmid LRIG-EGFP, LRIG3 fusion protein was found in kytoplasm, and the number of cells in the S phase increase and the ability of cell invasion descend obviously. The expression of EGFR and LRIG3 are different in different times after intervention with 100ng/ml EGF, the expression of EGFR decrease while the expression of LRIG3 increase as time extension.Conclusion The biology activity of LRIG3 is achieved through EGFR signal transduction pathway. It can inhibit the proliferation and the invasiveness of glioblastoma. LRIG3 gene may be one of the new key gene in gene therapy of glioblastoma.