Analysis of p53 codon 72 gene polymorphism in  Isfahanian patients with endometriosisDr. Mehdi Nikbakht Dastjerdi (PhD)

Associate professor, Department of Anatomical Sciences and Molecular Biology ,Faculty of Medicine, Isfahan University of Medical Sciences

Bahram Eslami Farsani(MSc) Senior Student, Faculty of Medicine, Isfahan University of Medical Sciences

.Roshanak Abutorabi(PhD)

Associate professor, Department of Anatomical Sciences and Molecular Biology ,Faculty of Medicine, Isfahan University of Medical Sciences

MethodsObjectives Results

Conclusions

References

The ages of samples were between 23-50 years old To analyze the codon 72 polymorphism, we used a PCR-based assay that specifically, amplify either TP53 Pro or TP53 Arg allele and give a PCR product by using specific primers for Arg allele (Figure 1) and/or Pro allele (Figure 2) respectively. Detection of TP53 codon 72 polymorphism by allele specific PCR was successfully conducted in all cases and controls. The distribution of the three different genotypes of codon 72 in exon 4 of TP53 in our cases and controls is shown in table 1.

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In this study, polymorphism of codon72 TP53 gene has been studied among 90 patients suffering endometriosis and 90 blood samples from normal people as control group. Different genotype of codon72 exon4 TP53 gene was determined by using allele specific polymerase chain reaction. the TP53 codon 72 Pro allele were detected by PCR using the primer pair p53Pro+/ p53Pro- (p53Pro+: 5′-GCCAGAGGCTGCTCCCCC; and P53Pro-: 5′-CGTGCAAGTCACAGACTT) and the p53 codon 72Arg allele by the primer pair p53Arg+/p53Arg-(p53Arg+: 5′-TCCCCCTTGCCGTCCCAA andp53Arg-: 5′CTGGTGCAGGGGCCACGC). The amplified products were subjected to electrophoresis on 1% agarose gel in 1× TBE buffer andvisualized on a transilluminator using ethidium bromide. Obtained information via software SPSS have been analyzed. For comparing the frequency distribution of three different genotype of codon 72 in endometriosis samples with that of control ones Chi-Square Test has been used. P-value lowers than 0.05% is considered meaningful.

Endometriosis is one of prevalent disease among women and has the same features of malignant tumors and its cause is not clear. Almost 10% women suffer this disease and nearly 18% women suffer during pregnancy age.This ratio reaches to nearly 30 to 40 % in infertile women .Endometriosis has been occurred by growing endometrial tissues outside the uterus and causes the abnormalities like painful sexual intercourse, painful menstruation and pelvic pains and infertility

Picture (1). Electrophoresis for Arginine allele

Picture (2). Electrophoresis gene for Proline allele

Genotype

Endometriosis Control group

Number Percent Number Percent

Arg/Arg 26 28.9 38 42.2

Pro/Pro 14 15.6 3 3.3

Arg/Pro 50 55.6 49 54.4

Table(1). Genotypic frequency distribution in control and endometriosis group

This study shows that genotype Pro/Pro codon72 exon4 P53 gene may be a predisposing genetic factor for endometriosis in Esfahan.