Linked Common Polymorphisms in the Gelatinase A Promoter Are Associated with Diminished Transcriptional Response to Estrogen and Genetic Fitness

Abstract

Gelatinase A (matrix metalloproteinase-2) plays a prominent role in multiple biologic processes. Prior studies have established critical roles for gelatinase A transcriptional regulation by defined enhancer elements. To determine possible functional single nucleotide polymorphisms within these elements, we determined the single nucleotide polymorphism distribution within 1,665 bp of the gelatinase A 5′-flanking region, using a healthy homogeneous Caucasian study group of 463 individuals. Among the polymorphisms detected, a G → A transition at bp –1575 was located immediately 5′ to a half-palindromic potential estrogen receptor binding site. In estrogen receptor-positive MCF-7 cells the –1575G allele functioned the allele the in binding of the estrogen to estrogen in estrogen estrogen receptor for distribution a → transition within binding at was in the –1575G → A studies of the allele the –1575G allele in of the –1575G → A transition the distribution in study of healthy of Caucasian a in Gelatinase A a the of a of polymorphism within estrogen receptor binding a in the biologic of polymorphism Gelatinase A (matrix metalloproteinase-2) plays a prominent role in multiple biologic processes. Prior studies have established critical roles for gelatinase A transcriptional regulation by defined enhancer elements. To determine possible functional single nucleotide polymorphisms within these elements, we determined the single nucleotide polymorphism distribution within 1,665 bp of the gelatinase A 5′-flanking region, using a healthy homogeneous Caucasian study group of 463 individuals. Among the polymorphisms detected, a G → A transition at bp –1575 was located immediately 5′ to a half-palindromic potential estrogen receptor binding site. In estrogen receptor-positive MCF-7 cells the –1575G allele functioned the allele the in binding of the estrogen to estrogen in estrogen estrogen receptor for distribution a → transition within binding at was in the –1575G → A studies of the allele the –1575G allele in of the –1575G → A transition the distribution in study of healthy of Caucasian a in Gelatinase A a the of a of polymorphism within estrogen receptor binding a in the biologic of polymorphism of the multiple roles in the of cells of in the of the gelatinase A a of In to the role of gelatinase A in in the of gelatinase have of the was to the of a single nucleotide polymorphism single nucleotide estrogen single nucleotide estrogen in the of the gelatinase A in a by of gelatinase A gelatinase A in of gelatinase A of multiple of of gelatinase A a of in a these have to the transcriptional the of gelatinase A studies have within the have within defined a binding for in the gelatinase A for gelatinase A in In for within the of cells defined a enhancer located bp to the transcriptional in the gelatinase A A in the gelatinase A was defined by studies have enhancer the in gelatinase A of the of the of gelatinase (matrix have the binding of polymorphism within the gelatinase transcriptional was of the role of gelatinase A in multiple the of the by the gelatinase we of the of the gelatinase A for polymorphisms transcriptional In we to the polymorphisms in the enhancer the binding the transcriptional regulation of gelatinase A in processes. the of the distribution of polymorphisms in the gelatinase A a functional at bp we polymorphisms within the enhancer region, a G → A transition was to bp –1575 was immediately to a potential half-palindromic estrogen receptor binding site. In we the distribution of the of the estrogen the –1575G → A to the → the transcriptional of gelatinase A to estrogen receptor binding a in of was in 463 healthy Caucasian using the in in for of Caucasian was within the of the Gelatinase A of was the of the bp to the transcriptional of the gelatinase A the the of determine the for the of of the gelatinase A by using the a bp to a bp to the allele in a in the –1575G the allele in a the of the for of for the polymorphism for the using To determine the of potential the the functional a of the of 463 was the bp to the –1575 bp MCF-7 the cells in cells for a to in to Gelatinase A of of the gelatinase A 5′-flanking was by the using the at bp to the transcriptional To the functional of the was To the of the polymorphism the was the allele of the polymorphism by was in the in of was by of bp to the the of the for the in to of MCF-7 cells to a in of was in at of the of to of the estrogen by estrogen to the estrogen of of in the of the these MCF-7 cells to at of in binding of of in a of for at the in a by in to the for at by of the for at of estrogen estrogen to of determined for using by of for multiple of for determined by using of within the of the Gelatinase A bp of the of the gelatinase A was to the of nucleotide within these study A bp to bp to a G → A transition at –1575 at polymorphism was a for the distribution studies defined binding site. the of the allele A in the allele the allele in a A was the –1575G allele the allele bp bp in of in bp bp in for the allele in of bp for the allele of the was the polymorphism the polymorphism by to determine potential the –1575 the polymorphism of the using the the the –1575 bp located immediately 5′ to a half-palindromic binding for the estrogen receptor a functional of the –1575 to determine the polymorphism to the half-palindromic gelatinase A in to of to the to the half-palindromic located bp was of the estrogen receptor was by the to the bp to in the gelatinase A G A at the –1575 the estrogen receptor-positive the estrogen In the of the MCF-7 the the G allele of a single of was MCF-7 the the A allele binding was the the estrogen cells the –1575G allele estrogen receptor-positive MCF-7 cells estrogen of the in a in using the –1575G the allele was using of MCF-7 binding to the was by A of –1575G for MCF-7 in the of the a of was a of the was the of MCF-7 the the –1575G allele MCF-7 of binding by of the –1575G allele the allele a of the was the MCF-7 a estrogen a estrogen the of the studies a of the estrogen in MCF-7 the half-palindromic of estrogen binding to the MCF-7 cells of the estrogen estrogen have a binding of at for the estrogen receptor a the the –1575G allele MCF-7 receptor was by the a of the Gelatinase A of of the gelatinase A 5′-flanking was to the by receptor-positive MCF-7 cells for studies the by a the a of to the of by using the of of a in was the the binding at A in was the estrogen receptor half-palindromic binding the a of by studies the of enhancer within the bp to the estrogen receptor half-palindromic binding of MCF-7 cells of the 5′-flanking of the gelatinase A a elements, the half-palindromic estrogen receptor binding the of the a of the of the functional of the polymorphism gelatinase A transcriptional the was by to of MCF-7 cells the of the of the the estrogen in the polymorphism at bp a of was these MCF-7 cells in bp of the of the gelatinase A the –1575G in the of MCF-7 cells the G A the G allele was a of the of at of the functional of the polymorphism within the half-palindromic estrogen receptor binding G A of of the the –1575 polymorphism in of a of MCF-7 cells the a in the In the of the the estrogen cells estrogen receptor for the enhancer of the for of enhancer of a the –1575 site. of the a the was the in to the of the a of of of estrogen cells the estrogen the the of these studies in of cells the the a of of the estrogen in the of in of the In the of in by the estrogen of the the was of of the in of the the the in estrogen receptor binding determined by by the –1575G → A the of estrogen the half-palindromic binding the functional of the –1575G → A In the functional binding → transition the gelatinase A to of in cells of a the estrogen in of the a of receptor have defined for of the of at binding of binding A of was to of these of to the gelatinase A within the of the –1575 of these studies in of the the in in the in the of in the the in the of in of by the was transition at –1575 the to the estrogen receptor by the of the of these the by in half-palindromic binding transcriptional in a in the the the estrogen of the of for these was was the in to the of the in the a of was for of the distribution of at bp –1575 within a group of 463 healthy Caucasian the distribution for the polymorphism was in the distribution for the polymorphism within the study was by the of for the polymorphism at bp –1575 the by of a of a of a of of the polymorphisms in the 5′-flanking of the gelatinase A in a To the of the transition to potential possible of a study group of of Caucasian was in the for the study group for the the the healthy of the for the by of in the of the polymorphism in study of Caucasian in a A of multiple Prior studies have defined critical in the 5′-flanking of the gelatinase A of defined of In study we a Caucasian for within the 5′-flanking of the gelatinase A the to potential within the of the of study the distribution within the gelatinase A of the 5′-flanking polymorphisms defined in the transition in In study the by detected, transition was of polymorphisms by in the gelatinase A in a of the gelatinase A In polymorphisms at the of for these of A in cells cells to gelatinase A by cells in of gelatinase A of gelatinase A in by a of 463 Caucasian polymorphisms within the defined enhancer binding of the immediately to the transition half-palindromic binding for the estrogen using a of estrogen receptor functional for the functional of the G → A transition at bp –1575 estrogen binding gelatinase A the estrogen estrogen receptor binding in to the estrogen to the by the polymorphism was functional within the of estrogen the estrogen the the of the polymorphism located within the the estrogen receptor defined the estrogen receptor binding of half-palindromic the binding of at for a the of the estrogen receptor for half-palindromic the of the immediately the –1575 receptor binding potential for the –1575G → A transition a in estrogen receptor binding the by –1575G → A transition was to to a → transition to binding to estrogen receptor the estrogen receptor the gelatinase A in a the in study we to determine by to allele was to within the of the the functional allele distribution a the a in of of the the for in the the allele of the homogeneous of the study of to the the distribution to in the by of a study of of Caucasian in of the in for the allele in a study of in study group was the to these the of the by in a of by A plays role for the of the gelatinase of the gelatinase in the transition in of estrogen we the in of the gelatinase A transcriptional regulation the to we have the gelatinase A a functional polymorphism at bp –1575 the transcriptional to estrogen of the gelatinase A to the of critical of the multiple roles in the of cells of in the of the gelatinase A a of In to the role of gelatinase A in in the of gelatinase have of the was to the of a single nucleotide polymorphism single nucleotide estrogen single nucleotide estrogen in the of the gelatinase A in a by of gelatinase A gelatinase A in of gelatinase A of multiple of of gelatinase A a of in a these have to the transcriptional the of gelatinase A studies have within the have within defined a binding for in the gelatinase A for gelatinase A in In for within the of cells defined a enhancer located bp to the transcriptional in the gelatinase A A in the gelatinase A was defined by studies have enhancer the in gelatinase A of the of the of gelatinase (matrix have the binding of polymorphism within the gelatinase transcriptional was of the role of gelatinase A in multiple the of the by the gelatinase we of the of the gelatinase A for polymorphisms transcriptional In we to the polymorphisms in the enhancer the binding the transcriptional regulation of gelatinase A in processes. the of the distribution of polymorphisms in the gelatinase A a functional at bp we polymorphisms within the enhancer region, a G → A transition was to bp –1575 was immediately to a potential half-palindromic estrogen receptor binding site. In we the distribution of the of the estrogen the –1575G → A to the → the transcriptional of gelatinase A to estrogen receptor binding a in of was in 463 healthy Caucasian using the in in for of Caucasian was within the of the Gelatinase A of was the of the bp to the transcriptional of the gelatinase A the the of determine the for the of of the gelatinase A by using the a bp to a bp to the allele in a in the –1575G the allele in a the of the for of for the polymorphism for the using To determine the of potential the the functional a of the of 463 was the bp to the –1575 bp MCF-7 the cells in cells for a to in to Gelatinase A of of the gelatinase A 5′-flanking was by the using the at bp to the transcriptional To the functional of the was To the of the polymorphism the was the allele of the polymorphism by was in the in of was by of bp to the the of the for the in to of MCF-7 cells to a in of was in at of the of to of the estrogen by estrogen to the estrogen of of in the of the these MCF-7 cells to at of in binding of of in a of for at the in a by in to the for at by of the for at of estrogen estrogen to of determined for using by of for multiple of for determined by using of was in 463 healthy Caucasian using the in in for of Caucasian was within the of the Gelatinase A of was the of the bp to the transcriptional of the gelatinase A the the of determine the for the of of the gelatinase A by using the a bp to a bp to the allele in a in the –1575G the allele in a the of the for of for the polymorphism for the using To determine the of potential the the functional a of the of 463 was the bp to the –1575 bp MCF-7 the cells in cells for a to in to Gelatinase A of of the gelatinase A 5′-flanking was by the using the at bp to the transcriptional To the functional of the was To the of the polymorphism the was the allele of the polymorphism by was in the in A of was by of bp to the the of the for the in to of MCF-7 cells to a in of was in at of the of to of the estrogen by estrogen to the estrogen of of in the of the these MCF-7 cells to at of in binding of of in a of for at the in a by in to the for at by of the for at of estrogen estrogen to of determined for using by of for multiple of for determined by using of within the of the Gelatinase A bp of the of the gelatinase A was to the of nucleotide within these study A bp to bp to a G → A transition at –1575 at polymorphism was a for the distribution studies defined binding site. the of the allele A in the allele the allele in a A was the –1575G allele the allele bp bp in of in bp bp in for the allele in of bp for the allele of the was the polymorphism the polymorphism by to determine potential the –1575 the polymorphism of the using the the the –1575 bp located immediately 5′ to a half-palindromic binding for the estrogen receptor a functional of the –1575 to determine the polymorphism to the half-palindromic gelatinase A in to of to the to the half-palindromic located bp was of the estrogen receptor was by the to the bp to in the gelatinase A G A at the –1575 the estrogen receptor-positive the estrogen In the of the MCF-7 the the G allele of a single of was MCF-7 the the A allele binding was the the estrogen cells of MCF-7 binding to the was by A of –1575G for MCF-7 in the of the a of was a of the was the of MCF-7 the the –1575G allele MCF-7 of binding by of the –1575G allele the allele a of the was the MCF-7 a estrogen a estrogen the of the studies a of the estrogen in MCF-7 the half-palindromic of estrogen binding to the MCF-7 cells of the estrogen estrogen have a binding of at for the estrogen receptor a the the –1575G allele MCF-7 receptor was by the a of the Gelatinase A of of the gelatinase A 5′-flanking was to the by receptor-positive MCF-7 cells for studies the by a the a of to the of by using the of of a in was the the binding at A in was the estrogen receptor half-palindromic binding the a of by studies the of enhancer within the bp to the estrogen receptor half-palindromic binding of MCF-7 cells of the 5′-flanking of the gelatinase A a elements, the half-palindromic estrogen receptor binding the of the a of the of the functional of the polymorphism gelatinase A transcriptional the was by to of MCF-7 cells the of the of the the estrogen in the polymorphism at bp a of was these MCF-7 cells in bp of the of the gelatinase A the –1575G in the of MCF-7 cells the G A the G allele was a of the of at of the functional of the polymorphism within the half-palindromic estrogen receptor binding G A of of the the –1575 polymorphism in of a of MCF-7 cells the a in the In the of the the estrogen cells estrogen receptor for the enhancer of the for of enhancer of a the –1575 site. of the a the was the in to the of the a of of of estrogen cells the estrogen the the of these studies in of cells the the a of of the estrogen in the of in of the In the of in by the estrogen of the the was of of the in of the the the in estrogen receptor binding determined by by the –1575G → A the of estrogen the half-palindromic binding the functional of the –1575G → A In the functional binding → transition the gelatinase A to of in cells of a the estrogen in of the a of receptor have defined for of the of at binding of binding A of was to of these of to the gelatinase A within the of the –1575 of these studies in of the the in in the in the of in the the in the of in of by the was transition at –1575 the to the estrogen receptor by the of the of these the by in half-palindromic binding transcriptional in a in the the the estrogen of the of for these was was the in to the of the in the a of was for of the distribution of at bp –1575 within a group of 463 healthy Caucasian the distribution for the polymorphism was in the distribution for the polymorphism within the study was by the of for the polymorphism at bp –1575 the by of a of a of a of of the polymorphisms in the 5′-flanking of the gelatinase A in a To the of the transition to potential possible of a study group of of Caucasian was in the for the study group for the the the healthy of the for the by of in the of the polymorphism in study of Caucasian in a of within the of the Gelatinase A bp of the of the gelatinase A was to the of nucleotide within these study A bp to bp to a G → A transition at –1575 at polymorphism was a for the distribution studies defined binding site. the of the allele A in the allele the allele in a A was the –1575G allele the allele bp bp in of in bp bp in for the allele in of bp for the allele A of the was the polymorphism the polymorphism by to determine potential the –1575 the polymorphism of the using the the the –1575 bp located immediately 5′ to a half-palindromic binding for the estrogen receptor a functional of the –1575 to determine the polymorphism to the half-palindromic gelatinase A in to of to the to the half-palindromic located bp was of the estrogen receptor was by the to the bp to in the gelatinase A G A at the –1575 the estrogen receptor-positive the estrogen In the of the MCF-7 the the G allele of a single of was MCF-7 the the A allele binding was the the estrogen cells of MCF-7 binding to the was by A of –1575G for MCF-7 in the of the a of was a of the was the of MCF-7 of the was the MCF-7 a estrogen a estrogen the of the studies a of the estrogen in MCF-7 the half-palindromic of estrogen binding to the MCF-7 cells of the estrogen estrogen have a binding of at for the estrogen receptor a of the Gelatinase A of of the gelatinase A 5′-flanking was to the by receptor-positive MCF-7 cells for studies the by a the a of to the of by using the of of a in was the the binding at A in was the estrogen receptor half-palindromic binding the a of by studies the of enhancer within the bp to the estrogen receptor half-palindromic binding To the functional of the polymorphism gelatinase A transcriptional the was by to of MCF-7 cells the of the of the the estrogen in the polymorphism at bp a of was these MCF-7 cells in of the functional of the polymorphism within the half-palindromic estrogen receptor binding G A of of the the –1575 polymorphism in of a of MCF-7 cells the a in the In the of the the estrogen cells estrogen receptor for the enhancer of the of of estrogen cells the estrogen the the of these studies in of cells the the a of of the estrogen in the of in of the In the of in by the estrogen of the the was of of the in of the the the in estrogen receptor binding determined by by the –1575G → A the of estrogen the half-palindromic binding the functional of the –1575G → A In the functional binding → transition the gelatinase A to receptor have defined for of the of at binding of binding A of was to of these of to the gelatinase A within the of the –1575 of these studies in of the the in in the in the of in the the in the of in of by the was transition at –1575 the to the estrogen receptor by the of the of these the by in half-palindromic binding transcriptional in a was for of the distribution of at bp –1575 within a group of 463 healthy Caucasian the distribution for the polymorphism was in the distribution for the polymorphism within the study was by the of for the polymorphism at bp –1575 the by of a of a of a of To the of the transition to potential possible of a study group of of Caucasian was in the for the study group for the the the healthy of the for the by of in the A of multiple Prior studies have defined critical in the 5′-flanking of the gelatinase A of defined of In study we a Caucasian for within the 5′-flanking of the gelatinase A the to potential within the of the of study the distribution within the gelatinase A of the 5′-flanking polymorphisms defined in the transition in In study the by detected, transition was of polymorphisms by in the gelatinase A in a of the gelatinase A In polymorphisms at the of for these of A in cells cells to gelatinase A by cells in of gelatinase A of gelatinase A in by a of 463 Caucasian polymorphisms within the defined enhancer binding of the immediately to the transition half-palindromic binding for the estrogen using a of estrogen receptor functional for the functional of the G → A transition at bp –1575 estrogen binding gelatinase A the estrogen estrogen receptor binding in to the estrogen to the by the polymorphism was functional within the of estrogen the estrogen the the of the polymorphism located within the the estrogen receptor defined the estrogen receptor binding of half-palindromic the binding of at for a the of the estrogen receptor for half-palindromic the of the immediately the –1575 receptor binding potential for the –1575G → A transition a in estrogen receptor binding the by –1575G → A transition was to to a → transition to binding to estrogen receptor the estrogen receptor the gelatinase A in a the in study we to determine by to allele was to within the of the the functional allele distribution a the a in of of the the for in the the allele of the homogeneous of the study of to the the distribution to in the by of a study of of Caucasian in of the in for the allele in a study of in study group was the to these the of the by in a of by A plays role for the of the gelatinase of the gelatinase in the transition in of estrogen we the in of the gelatinase A transcriptional regulation the to we have the gelatinase A a functional polymorphism at bp –1575 the transcriptional to estrogen of the gelatinase A to the of critical Gelatinase A of multiple Prior studies have defined critical in the 5′-flanking of the gelatinase A of defined of In study we a Caucasian for within the 5′-flanking of the gelatinase A the to potential within the of the of study the distribution within the gelatinase A of the 5′-flanking polymorphisms defined in the transition in In study the by detected, transition was of polymorphisms by in the gelatinase A in a of the gelatinase A In polymorphisms at the of for these of Gelatinase A in cells cells to gelatinase A by cells in of gelatinase A of gelatinase A in by a of 463 Caucasian polymorphisms within the defined enhancer binding of the immediately to the transition half-palindromic binding for the estrogen using a of estrogen receptor functional for the functional of the G → A transition at bp –1575 estrogen binding gelatinase A the estrogen estrogen receptor binding in to the estrogen to the by the polymorphism was functional within the of estrogen the estrogen the the of the polymorphism located within the the estrogen receptor defined the estrogen receptor binding of half-palindromic the binding of at for a the of the estrogen receptor for half-palindromic the of the immediately the –1575 receptor binding potential for the –1575G → A transition a in estrogen receptor binding the by –1575G → A transition was to to a → transition to binding to estrogen receptor the estrogen receptor the gelatinase A in a the in study we to determine by to allele was to within the of the the functional allele distribution a the a in of of the the for in the the allele of the homogeneous of the study of to the the distribution to in the by of a study of of Caucasian in of the in for the allele in a study of in study group was the to these the of the by in a of by Gelatinase A plays role for the of the gelatinase of the gelatinase in the transition in of estrogen we the in of the gelatinase A transcriptional regulation the to In we have the gelatinase A a functional polymorphism at bp –1575 the transcriptional to estrogen of the gelatinase A to the of critical