Paper Title
Reference Gene Identification in Blood Samples of Carcinoma Lung Patients Using Quantitative Real-Time PCR

Abstract
Background: Several techniques of assay for early detection of cancer that improves the survival rates has been employed in tissues and cell lines. RTqPCR is one of the most common technique employed for gene expression studies for normalization of target gene using reference gene (RG). Present study used three most common RGs: GAPDH, β-Actin and 18s rRNA which were assessed by qPCR to validate as of which is more effective reference gene in blood samples of lung cancer patients. Materials and Methods: Total 30 subjects with Lung Cancer of Non-Small Cell and Small Cell were included along with 20 healthy controls. RNA isolated from peripheral blood mononuclear cells was quantified, prepared for cDNA synthesis and analyzed for expression of three reference genes on RTqPCR. Results: Expression levels as Ct values of studied reference genes were reported as Mean±SD for GAPDH (26.97±5.107), β-actin (20.5±2.3) and 18s rRNA (25.10±4.075). GAPDH showed lowest expression whereas β-Actin showed highest expression among the studied reference genes in subjects of Lung Cancer. The expression of GAPDH and 18s were statistically significantly lower than β -actin (p<0.0001), whereas expression levels of GAPDH and 18s rRNA were comparable. However expression level of only β-actin in Lung Cancer subjects was comparable with healthy controls with p<0.1611 at 95% CI. Conclusion: It is concluded that β -Actin to be considered as most suitable reference gene isolated and studied from peripheral blood mononuclear cells using RT qPCR in Lung Cancers. Acknowledgment: This study has been supported by the Department of Science and Technology New Delhi under WOS-A Project (SR/WOS-A/LS-605/2016(G) Keywords - Reference Gene, Carcinoma Lung, Gene Expression