Detection of Methylene Tertahydrofolate Reductase C677T Mutation among Sudanese Patients with Prostate Cancer

Abstract

A common 677 C-T transition (rs1801133) in the MTHFR gene is a well identified genetic determinant of hyperhomocysteinemia and there are some reports have shown an association between MTHFR gene polymorphism with cancer development. The aim of this study was to detect the presence of MTHFR polymorphism among Sudanese prostate cancer patients by using PCR and explore its relation with hypercoagulable state. The study is a case control study conducted at Taiba cancer center and Khartoum center for oncology in period from June to December 2017, 38 patients with prostate cancer(diagnosed by histopathology) and 40 healthy male(control group) were enrolled in this study,2.5 ml venous blood was collected after informed consent. RBCS was Hemolysed by alkaline solution (Red Cells lysis buffer) ,then the membranes of WBC were digested by solution containing detergent and proteases (White Cells Lysis buffer),then protein was precipitated out by saturated NaCL and centrifugation , finally DNA was precipitated by absolute ethanol ,washed by 70% ethanol and eluted in 50 μL of 10 mM Tris-HCl, 1 mM EDTA, pH 8.0 . MTHFR C677T genotype frequencies were detected by PCR, Five μl of the PCR product (ready to load) was electrophoresed on 1.5% agarose gel, and was stained with ethedium bromide, 1X TBE buffer was used as a running buffer. The Voltage applied to the gel was 100 volt with time duration of 30 minutes. 50 bp DNA ladder was used as molecular weight marker with each patch of samples .Finally, PCR product was demonstrated by gel system.The frequencies of CC and TT genotypes among the patients with prostate cancer were 95 % and 5% respectively, and among the control subjects 97.5 %, and 2.5%, respectively. In conclusion, there was no statistically significant difference in genotypes distribution when compered in patients with prostate cancer and control so thrombosis for those patients not caused by MTHFR gene mutation