Genes dysregulated significantly in tumor tissues compared with their normal counterparts are always considered as biomarkers or closely associated with carcinogenesis. Over the past two decades plentiful efforts have
been devoted to the identification of genes selleck compound involved in cancer development [1]. Many approaches have been used to compare gene expression between two different physiological states. Differential Display (DD) is a useful method to compare patterns of gene expression in RNA samples of different types or under different biological conditions [2, 3]. The technique produces partial cDNA fragments by a combination of reverse transcription and PCR of randomly primed RNA. Changes in the expression level of genes are identified after separation of the
cDNA fragments produced in an arbitrarily primed polymerase chain reaction on a sequencing-type gel. Combined with RNA expression verification, Differential Display is a powerful method for generating high confidence hits in the screening of hundreds of potential differentially expressed transcripts. Lung cancer is one of the most common human cancers and the leading cause of cancer death worldwide [4, 5]. With the same genetic backgrounds but different metastatic buy AZD5582 potential, 95C and 95D cell lines were subcloned from a poorly differentiated human large cell lung carcinoma cell line PLA-801 by Dr. Lezhen Chen (Department of Pathology, Chinese PLA General Hospital), which were suitable for Differential Display analysis. Nude mice incubated
with 95D cells showed earlier and more metastasis than incubated with 95C cells [6, 7]. Although the importance of tumorigenesis has been realized and studied, limited knowledge is known about its associated genes and signal networks. Understanding Glycogen branching enzyme further more players and intrinsic processes involved in carcinogenesis could lead to effective, targeted strategies to prevent and treat cancer. In the present study, we found that LCMR1 was expressed significantly higher in 95D cell line compared to 95C using a combination of DD-PCR and real-time PCR. We then investigated its expression in various human tissues by northern blot. Recombinant LCMR1 protein was expressed and its specific polyclonal antibody was generated. To examine its involvement in carcinogenesis, 84 specimens of NSCLC patients were examined for the expression of LCMR1 by immunohistochemistry analysis. Our results strongly suggested that LCMR1 was significantly overexpressed in human NSCLC and its expression was closely associated with clinical stage of patients with NSCLC, which may have applications in lung cancer diagnosis and treatment. Materials and methods Cell lines 95C and 95D cell lines were subcloned from a poorly differentiated human large cell lung carcinoma cell line PLA-801 and kindly provided by Dr. Lezhen Chen (Department of Pathology, Chinese PLA General Hospital, China).