SNPs in the Regulatory Region of p53 Gene in Women with Gynaecological Diseases

• Main Authors: Ching-Wei Huang, 黃清瑋
• Other Authors: Chich-Sheng Lin
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/89988809448408288537

碩士 === 國立交通大學 === 生物科技研究所 === 91 === Objective: To investigate the genotypes of progesterone receptor gene（PROGINS ）, p53 coding region, and upstream of p53 gene from women with gynaecological diseases such as endometriosis, uterine leimyoma, adenomyosis, ovarian cancer, polycystic ovary syndrome (PCOD), and hyperprolactinemia. Background: Through the study of gene, scientists could understand the mechanism and probability of diseases, especial the studies of p53. About 55% of human cancers suffer mutations in both alleles of p53 gene. The high association between p53 mutations and many forms of cancers suggests that loss of p53 function can be used as a marker for high susceptibility to carcinogenesis. So far, genetic polymorphism of p53 gene regulatory region associated with carcinogenesis is unclear. Therefore, to find out the polymorphism of p53 regulatory region which related with gynaecological diseases remains to be explored. Materials and Methods: A total of 196 blood samples, including 43 uterine leiomyoma, 15 adenomyosis, 23 endometriosis, 22 ovarian cancer, 15 PCOD, 23 hyperprolactinemia, and 43 healthy individuals were collected and genomic DNA were prepared. Determination of PROGINS was performed by PCR and gel electrophoresis. Determination of p53 coding region codon 11 and 248 were performed by PCR and restriction fragment length polymorphism（RFLP）. The p53 regulatory region (-433 ~ +29) was amplified by PCR and then the sequence was idnetified. To find out the single nucleotide polymorphisms (SNPs) in the p53 regulatory region, the multiple sequences were aligned with program “Clustal W”. The SNPs between patients and healthy individuals were evaluated by chi-square test of independence and odds ratio. Results: A total of 154 samples, including 41 healthy individuals, of PROGINS and p53 coding region codon 11 and 248 were studied. Frequencies of T1 (159 bp) allele was 98.4% and T2 (480 bp) allele was 1.6% in the both populations of patients and healthy individuals. Morever, no mutant types of p53 codon 11 and 248 were discovered in the both populations. Therefore, there were no statistically significant association between PROGINS, p53 codon 11 and 248 from women with gynaecological diseases to the normal. In the analysis of p53 regulatory region, 140 sequences, including 43 healthy individuals have been sequenced. 98 mutations and 15 SNPs have been identified in the p53 regulatory region. There was no statistically significant association among the SNPs from the gynaecological diseases compared with the normal. However, the SNP of -51 T/A, on the binding site of NF-B and in the response region of genotoxic stress, was happened in the patients of unterine leiomyomas and healthy individuals that the prevalence rate was up to 10% in the unterine leiomyomas. The SNP -33 A/G, on the c-Myc/Max heterodimer binding site, was only happened in the unterine leiomyomas. The result hints that though the p value is greater than 0.05, the relationship between the SNPs and gynaecological diseases need to be more discussions. Conclusion: This study is a pioneer to report the SNPs in the regulatory region of p53 gene. The results in the present study provide the capacity to study the relationship between gene polymorphism in the regulatory region and diseases in a large scale. We wonder to expect the critical results concerning with gynaecological diseases associated with the p53 SNPs will be published in the future.