International Journal of Medical Laboratory Research (IJMLR)

IMMUNOHISTOCHEMISTRY OPTIMISATION OF SOME TUMOUR SUPPRESSOR GENES IN CHRONIC OBSTRUCTIVE PULMONARY DISEASE ON FORMALIN FIXED PARAFFIN EMBEDDED TISSUE SECTIONS

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Davou GI, Ekwempu AI, BotD, Chuwang NJ, Adisa JO

 

ABSTRACT: Background: Lung diseases has been shown to be among the most occurring diseases globally with diseases ranging from self-limiting flues to life threatening chronic diseases such as chronic obstructive pulmonary disease (COPD) and lung malignancies. Tobacco has been identified as the major singular risk factor for the pathogenesis of chronic pulmonary diseases. The incidence of primary lung carcinoma (PLC) is increased in COPD Patients due to an existing impairment in normal lung function. This study was aimed at carrying out preliminary IHC optimisation for p53, TTC5, Bax and p21 tumour suppression genes on COPD tissue sections. Method: Several IHC staining using the avidin- biotin complex kit (ABC) were carried out to express the four genes using different staining conditions and different primary antibody dilutions. Results: Our study revealed high-level expression of the four tumour suppression genes on COPD tissue sections with 1:250, 1:500 1: 500 and 1:100 optimum primary antibody dilutions for p53, TTC5, Bax and p21 genes respectively. All experiments for optimum results were carried out under 21⁰C room temperature. Conclusions: These findings revealed an active DNA -8damage response, even though we are still far from understanding the current function of p53 and its downstream genes in this particular tissue. Further investigations are required and the application of more techniques for an expanded study with more access to lung cancer tissue sections from different cohorts, in order to fully understand the role of p53 and the other genes that respond to DNA damage in lung cancer therapy.

 

KEY WORDS: IHC, Optimisation, COPD, Lung cancer, DNA damage response, P53, Laboratory research

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