Altered Secretory Activity of APE1/Ref-1 D148E Variants Identified in Human Patients With Bladder Cancer.
- Author:
Yu Ran LEE
1
;
Jae Sung LIM
;
Ju Hyun SHIN
;
Sunga CHOI
;
Hee Kyoung JOO
;
Byeong Hwa JEON
Author Information
1. Department of Physiology, Chungnam National University School of Medicine, Daejeon, Korea. bhjeon@cnu.ac.kr
- Publication Type:Original Article
- Keywords:
Apurinic/Apyrimidinic Endonuclease 1/Redox Factor-1 (APE1/Ref-1);
Point Mutation;
Secretion;
Enzyme-Linked Immunosorbent Assay;
Bladder Cancer
- MeSH:
Base Sequence;
Clinical Coding;
DNA Repair;
Enzyme-Linked Immunosorbent Assay;
Genetic Variation;
HEK293 Cells;
Humans*;
Oxidation-Reduction;
Point Mutation;
Polymerase Chain Reaction;
Reverse Transcription;
Sequence Analysis, DNA;
Urinary Bladder Neoplasms*;
Urinary Bladder*
- From:International Neurourology Journal
2016;20(Suppl 1):S30-S37
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein involved in DNA repair and redox modulation. Recently, serum and urinary APE1/Ref-1 levels were reported to be increased in patients with bladder cancer. Genetic variations of APE/Ref-1 are associated with the risk of cancer. However, the effect of APE1/Ref-1 variants on its secretory activity is yet unknown. METHODS: APE1/Ref-1 variants were evaluated by DNA sequencing analysis of reverse transcription polymerase chain reaction products in coding DNA sequences (CDS) of APE1/Ref-1 in bladder tissue samples from patients with bladder cancer (n=10). Secretory activity of APE1/Ref-1 variants was evaluated with immunoblot and enzyme-linked immunosorbent assay of the culture medium supernatants. RESULTS: Four different substitution mutants (D148E, I64V/D148E, W67R/D148E, and E86G/D148E) of APE1/Ref-1 were identified in bladder cancer specimens. However, deletion mutants of APE1/Ref-1 CDS were not found. The secretory activity of the APE1/Ref-1 variants (D148E, I64V/D148E, and E86G/D148E) was increased compared to that of wild type APE1/Ref-1. Furthermore, the secretory activity in basal or hyperacetylated conditions was much higher than that in APE1/Ref-1 D148E-transfected HEK293 cells. CONCLUSIONS: Taken together, our data suggest that the increased secretory activity of D148E might contribute to increased serum levels of APE1/Ref-1 in patients with bladder cancer.
