The Relation between Survivin Gene Expression and Urinary Bladder Cancer Disease

Abstract

Purpose: The aim of this study is to investigate the expression of survivin gene in patients suffering from urinary bladder cancer, thus contributing to further understanding of the molecular etiology of the disease by assessment of Survivin gene expression on both RNA and protein levels in urinary bladder tissue samples and Correlate between its expression and pathological prognostic parameters of patients with urinary bladder cancer risk. Methods:In the present study 26 cases of urinary bladder cancer and 26 cases of normal tissues far from tumor sites in the urinary bladder of the same patients (as control samples) were collected, also 10 paraffin-embedded tissue samples from consecutive of archival bladder specimens patients and 10 tissue samples that of healthy tissue from the same individual as control group were collected for immunohistochemical detection of survivin protein and real time PCR for survivin mRNA detection. Results:In normal tissues of control samples no expression for survivin gene has been noticed. However, in patients affected with bladder cancer the survivin expression was highly increased by increasing degree of tumor malignancy as indicated by real time PCR for survivin mRNA and immunohistochemistry for survivin protein. Conclusion:From this study, it is concluded that survivin protein was not expressed in normal bladder urothelium but was present in a high percentage in tissue samples of patients suffering from bladder cancer and its expression is associated with disease recurrence, tumor grade, progression and mortality. Hospice and Palliative Medicine International Journal Research Article Open AccessThe relation between survivin gene expression and urinary bladder cancer disease ii. Samples that used to detect mRNA expression levels: Tissue samples were taken from tumor masses from urinary bladder tissues (26 tumor samples) and normal tissues far from tumor sites in the urinary bladder as control samples (26 control samples from the same patients) . All samples were stored at (-80°C) until they are used.iii. Samples that used for protein expression detection: 10 paraffin-embedded tissue samples from consecutive of archival bladder specimens patients, also 10 tissue samples that of healthy tissues from the same individuals as control group.Immunohistochemical staining for survivin was done using the procedure (According to: Buchwalow and Bocher, 2010) and Immunohistochemical staining protocol (Genemed, USA):The list of reagents used for Immunohistochemical staining and mRNA extraction process could be found in (Table 1). ii. Different concentration of ethyl alcohol (30%, 50%, 70%, 80%, 90%, 95% and100%).iii. Xylene .iv. Paraffin wax. ii. Survivin reverse primer : 5΄CAA CCG GAC GAA TGC TTT T 3΄iii. Survivin probe: 5΄ CCA GAT GAC GAC CCC ATA GAG GAA CA 3΄ labeled by VIC® as a reporter dye on the 5´ end and TAMRA™ as a quencher dye on the 3´ end.iv. TaqMan Universal PCR Master Mix (Applied Biosystems, USA).The procedure included i. Fixation of tissues in 10% neutral buffered fo...