Author(s): Thangam Ramar | Kannan Soundarapandian | Suresh Babu | Gunasekaran Palani | Kaveri Krishnasamy | Mohana Sambasivam | Kavita Arunagiri | Dhanagaran Devaraj | Raja Sundaravadivel
Journal: International Journal of Biomedical Research
ISSN 0976-9633
Volume: 2;
Issue: 5;
Start page: 320;
Date: 2011;
Original page
Keywords: Cyclooxygenase 2 | cDNA quantification | Carcinoma | Reverse Transcription | PCR | Real Time -PCR
ABSTRACT
In advanced studies Cyclooxygenase-2 (COX-2) mRNA was determined mostly by in situ hybridization or Northern Blot analysis-methods not suitable for absolute quantification of mRNA copy numbers. Here we reported that, over expression of COX-2 mRNA was observed and calibrated through highly sensitive externally standardized real-time RT-PCR with Light Cycler instrument. The total RNA was isolated from two breast cancer cell lines (MCF-7, ZR-75-1), one lung cancer cell line (A549) and one normal breast cell line (HBL-100). The presence of COX-2 mRNA copy numbers was determined in all cell lines thourgh Reverse Transcription PCR by cDNA conversion method. In this study, we observed breast cancer cell lines and lung cancer cell lines were showing high expression levels of COX-2 mRNA at the same time there was the lowest expression was detected in normal breast cells. This low levels of COX 2 expression due to the tumorogenic action of a normal cells. Thus we evaluated COX-2 expression at different levels in breast and lung carcinoma cell lines. Results of our study provide insight view to the involvement of different carcinoma cells in pathogenesis with respect to COX-2 mRNA expression. This Light Cycler technology is currently considered to be the most precise method for nucleic acid quantification and which showed to be a powerful tool for further expression studies on cancer gene pathogenesis.
Journal: International Journal of Biomedical Research
ISSN 0976-9633
Volume: 2;
Issue: 5;
Start page: 320;
Date: 2011;
Original page
Keywords: Cyclooxygenase 2 | cDNA quantification | Carcinoma | Reverse Transcription | PCR | Real Time -PCR
ABSTRACT
In advanced studies Cyclooxygenase-2 (COX-2) mRNA was determined mostly by in situ hybridization or Northern Blot analysis-methods not suitable for absolute quantification of mRNA copy numbers. Here we reported that, over expression of COX-2 mRNA was observed and calibrated through highly sensitive externally standardized real-time RT-PCR with Light Cycler instrument. The total RNA was isolated from two breast cancer cell lines (MCF-7, ZR-75-1), one lung cancer cell line (A549) and one normal breast cell line (HBL-100). The presence of COX-2 mRNA copy numbers was determined in all cell lines thourgh Reverse Transcription PCR by cDNA conversion method. In this study, we observed breast cancer cell lines and lung cancer cell lines were showing high expression levels of COX-2 mRNA at the same time there was the lowest expression was detected in normal breast cells. This low levels of COX 2 expression due to the tumorogenic action of a normal cells. Thus we evaluated COX-2 expression at different levels in breast and lung carcinoma cell lines. Results of our study provide insight view to the involvement of different carcinoma cells in pathogenesis with respect to COX-2 mRNA expression. This Light Cycler technology is currently considered to be the most precise method for nucleic acid quantification and which showed to be a powerful tool for further expression studies on cancer gene pathogenesis.