Cloning, Sequence Analysis, Expression Study And Primary Function Determination Of A Novel Lung Cancer Metastasis Related Gene 1

Objective: The aim of the present study was to clone novel genes related to metastasis of lung cancer through differential display reverse transcript PCR(DDRT-PCR), which was performed between high-rate and low-rate metastasis large-cell lung cancer cell lines.Methodology: DDRT-PCR was performed in two large-cell lung cancer cell lines with different metastasis ability using HIEROGLYPH mRNA Profile System(Genomyx,Foster,Calif). Differentially expressed cDNAs were cloned and sequenced, searched against GenBank database. Beginning from a suitable EST, the full-length cDNA was obtained via RACE. Northern blotting was utilized to reveal the expression profile of the novel gene. Fusion protein of GFP and the new gene was constructed, transfected into human bronchial cells, then subcellular localization be revealed under fluro-microscopy. Further function investigation was encouraged after prokaryotic expression system was constructed and almost pure product was obtained.Results: We obtained 50 differentially displayed cDNAs, including 18 function-known genes, 16 function-unknown genes, 16 genomic DNA fragments and 4 ESTs. A novel full-length cDNA was gained through RACE, submitted to GenBank and nominated LCMR1. High expression of the gene was observed in human heart, kidney, muscles et al. The products of LCMR1 was localized in the cellular plasma. The prokaryotic expression system of LCMR1 was constructed, and conditions were optimized to reach a higher expression lever of LCMR1 product.