Discovery of Differentially Overexpressed Genes in Immortalized Cells and Human Pulmonary Non-small Cell Carcinomas

ABSTRACT

PURPOSE: Our aim of research is to find novel genes that overexpressed in various samples such as cell lines and tissues that infinitely proliferate; so-called immortalized cells, cancer cells and tissues. In this study, we obtained a gene from immortalized cell line (WI-38 VA13) then identified it from various cell lines and human lung tissues. MATERIALS AND METHODS: Using suppressive subtractive hybridization (SSH) method, we obtained many genes and selected a novel gene of them. And then the novel gene fragment was amplified by PCR and ligated in T easy vector for sequencing. And finally we found a differentially expressed gene in cell lines and tissues when it was performed by reverse transcriptase-PCR (RT-PCR). RESULTS: As the result of transformation of genes that we gained using SSH, we obtained about 150 clones. And then we certificated several genes by DNA prep and confirmed it by sequencing. We examined whether the gene sequences are novel or known genes by genome homology search and we confirmed the gene expressions by RT-PCR. As a result, we identified a differentially overexpressed gene (named "clone 58") in immortalized cells, cancer cell lines and lung squamous cell carcinomas. CONCLUSION: The "clone 58" mRNA was significantly up-regulated in various human cell lines and also human lung cancer tissues compared to the normal. We suppose that this gene can carry out a specific role related to the induction of cancer and/or the mechanism of the changeover of a normal cell to an immortalized cell. In short, the discovery of our gene has an importance in the point that they are thought to have a connection with immortalization and carcinogenesis of human cells and tissues.