American Journal of Biochemistry and Biotechnology

Detection and Localization of Anti and Pro-apoptotic mRNA Genes in Human Colorectal Cancer Using in situ RT-PCR

Mohd. Nazil Salleh, Siaw Shi Boon, Wan Khairuzzaman Wan Ramli, Azlina Zahari, Norashikin Shamsudin and Thuaibah Hashim

DOI : 10.3844/ajbbsp.2010.164.171

American Journal of Biochemistry and Biotechnology

Volume 6, Issue 3

Pages 164-171

Abstract

Problem statement: Recent studies using conventional Polymerase Chain Reaction have shown that anti-apoptotic (Cyclooxygenase-2, COX-2 and Nuclear Factor-κB, NF-κB) and proapoptotic mRNA (Bax and Bad) are involved in the tumorigenesis of colorectal cancer. The aim of this study was to localize the expression of anti and pro-apoptotic mRNA genes using Reverse Transcription in situ Polymerase Chain Reaction (RT in situ PCR) and immunodetection technique in the early stage of human colorectal adenocarcinoma. Approach: Reverse Transcription in situ Polymerase Chain Reaction (RT in situ PCR) and immunodetection technique was applied throughout of this studies. 20 paraffin-embedded tissue blocks of human colorectal adenocarcinoma samples was used compared to controls. Results: Morphologically, the glands and crypts were well-differentiated, enlarged and irregular with active secretion of mucin. COX-2, NF-κB, Bax and Bad mRNA were expressed in both normal and human colorectal cancer tissues. All mRNA genes were expressed in the cytoplasm and nuclei. However, COX-2 and NF-κB mRNA genes were highly expressed with higher intensity of brown staining compared to Bax and Bad at tubular epithelium cells. Conclusion: This study demonstrated that by using RT in situ PCR, COX-2 and NF-κB mRNA genes were shown to be involved in the development of human colorectal cancer.

Copyright

© 2010 Mohd. Nazil Salleh, Siaw Shi Boon, Wan Khairuzzaman Wan Ramli, Azlina Zahari, Norashikin Shamsudin and Thuaibah Hashim. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.