Assessing the expression and the increased expression of p21 gene in paraffin-embedded blocks of patients with bladder cancer who have been exposed to mitomycin


Authors:

Arsalan Jalili1 Sahar Shakeri Yekta1 Amin Ebrahimi Sadrabadi2 Elham roshandel3 1.Department of Biology School of Basic Science, Islamic Azad University, East Tehran branch, Tehran Iran 2.Department of Biology, Faculty of science, Islamic Azad university, Tonekabon branch, Tonekabon, Iran 3.Hematology Department, Facuilty of Medical Science, Shahid Beheshti University, Tehran, Iran

Correspondence:

Elham Roshandel - elham.roshandel@gmail.com

Background: bladder cancer arises from the epithelial lining layer of the urinary bladder. Understanding the molecular pathway of that can provide some useful informations about therapeutic manners. Hyper activity of P21 gene has been reported in recent years. The purpose of this study was to assess the expression and the increased expression of p21 gene in paraffin-embedded blocks of patients with bladder cancer before and after exposure to mitomycin.Materials and methods:in this study, 20 patients were selected and the blocks of their paraffin-embedded samples were prepared before and after exposing to mitomycin. After sectioning and paraffin removal, RNA was extracted and then cDNA synthesis was performed by using MMULV enzyme, Oligo dt and random hexamer primers. P21 specific primers and β-actin (as an internal control) were extracted from high cited articles. RT-PCR and Real Time PCR were performed for evaluation the expression and reduced expression of p21 gene level in samples exposed with mitomycin.Results: Real Time PCR showed the increased expression of 21 in samples which were exposed to mitomycin. Conclusion: the results suggest the lower expression of p21 gene in patient with bladder cancer and also the amount of p21 expression was increased in patients which were exposed by mitomycin. Investigating on increased expression of p21 gene in treated patients can be considered as a proper manner in researches on bladder cancer.Keyword:bladder cancer, p21, RT-PCR, Real Time PCR.