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Establish an In Vitro Model for the Study of NF2 Gene Function and Gene Therapy

机译:建立体外研究NF2基因功能和基因治疗的模型

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Two vestibular schwannoma cell lines, HEl-193 and HEl-182, as well as one vestibular nerve schwann cell line, HEl-286, were established using retroviral-mediated transfer of the HPV E6-E7 genes. HEI-193 cells, have a unique splice site mutation of the NF2 gene, while the NF2 gene in HEl-182 cells does not appear to be mutated. HEI-286 cells are expected to have the wildtype NF2 gene. Both immunocytochemistry and molecular biology techniques were used to demonstrate that these cell line are of Schwann cell origin. Comparison of the primary tumor with HEl-193 revealed the same NF2 mutation and an identical pattern of allele loss at multiple loci, indicating that the established cell line had maintained many of the properties of the original tumor. The immortalized cell lines were non- tumorigenic in severe combined immunodeficient (SCID) mice and/or nude mice, but did show altered growth properties such as higher proliferation rate and independence of Schwann cell growth factors. As part of our analysis, we also performed microarray analysis of gene expression in the cell lines as well as in tumors, including the original 193 tumor. The results obtained to date will help in our future studies aimed at elucidating the molecular pathogenesis of NF2.

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