Limits...
Significance of rs1271572 in the estrogen receptor beta gene promoter and its correlation with breast cancer in a southwestern Chinese population.

Chen L, Liang Y, Qiu J, Zhang L, Chen X, Luo X, Jiang J - J. Biomed. Sci. (2013)

Bottom Line: The rs1271572 G→T SNP abrogated YY1 binding and reduced the transcription activity of the promoter 0 N in the ERβ gene in vitro.TT genotype of rs1271572 inhibited expression of ERβ gene by down regulating transcriptional activity of the promoter 0 N in the ERβ gene.Our data revealed that the TT genotype of rs1271572 resulted in loss of the YY1 binding site and reduced the transcription activity of the promoter 0 N in the ERβ gene.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Background: To characterize single nucleotide polymorphisms (SNPs) within the promoter region of the estrogen receptor beta (ERβ) gene and to analyze the association of ERβ SNPs with susceptibility to breast cancer. Genotype frequencies of five SNPs (rs3020449, rs3020450, rs2987983, rs1271572 and rs1887994) in the promoter region of the ERβ gene in 873 women with breast cancer, 645 women with fibroadenoma and 700 healthy women were determined using an allele-specific tetra-primer polymerase chain reaction (PCR). Kaplan-Meier survival analysis was performed to evaluate the association of selected rs1271572 with prognosis in breast cancer. Electrophoretic mobility-shift assays were conducted to explore the binding of SNP rs1271572 containing probes to transcriptional factor Ying Yang 1 (YY1).

Results: Women with the homozygous TT genotype of rs1271572 had a significantly higher risk in developing breast cancer. Breast cancer patients with the TT genotype of rs1271572 had lower five-year survival rates than those with other genotypes and were more likely to suffer brain metastases. The rs1271572 G→T SNP abrogated YY1 binding and reduced the transcription activity of the promoter 0 N in the ERβ gene in vitro.

Conclusions: TT genotype of rs1271572 is associated with increased risk for breast cancer in Chinese women and is associated with unfavored prognosis in Chinese breast cancer patients. TT genotype of rs1271572 inhibited expression of ERβ gene by down regulating transcriptional activity of the promoter 0 N in the ERβ gene. Our data revealed that the TT genotype of rs1271572 resulted in loss of the YY1 binding site and reduced the transcription activity of the promoter 0 N in the ERβ gene.

Show MeSH

Related in: MedlinePlus

Binding of nuclear proteins in breast cancer cells to32 P- rs1271572G DNA probes. Different doses of nuclear proteins isolated from MDA-MB-231 cells were incubated with 32P-rs1271572G probes and the DNA-protein complexes were shown following autoradiography (lanes 1–7). The nuclear proteins isolated from MCF-7, MDA-MB-468, BT-549 and ZR75-30 were incubated with 32P -rs1271572G and the DNA-protein complexes were shown (lane 9–12). When 32Prs1271572T probes were used, no specific DNA-protein binding was observed (lanes 13 and 14). Binding activity of MDA-MB-231 nuclear extracts with 32P-rs1271572G probe was inhibited by cold rs1271572G probes(×50 times higher than 32P labeled probes) (lane 15). The DNA-protein complex was supershifted after further incubating with anti-YY1 antibody in MDA-MB-231 cells (lane 16). Position A indicated the binding complex in the presence of the anti-YY1 antibody. Position B indicated the binding complex of 32Prs1271572G DNA probes with nuclear proteins from the different breast cell lines. Position C indicated free 32P- rs1271572G DNA Probes.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3672062&req=5

Figure 5: Binding of nuclear proteins in breast cancer cells to32 P- rs1271572G DNA probes. Different doses of nuclear proteins isolated from MDA-MB-231 cells were incubated with 32P-rs1271572G probes and the DNA-protein complexes were shown following autoradiography (lanes 1–7). The nuclear proteins isolated from MCF-7, MDA-MB-468, BT-549 and ZR75-30 were incubated with 32P -rs1271572G and the DNA-protein complexes were shown (lane 9–12). When 32Prs1271572T probes were used, no specific DNA-protein binding was observed (lanes 13 and 14). Binding activity of MDA-MB-231 nuclear extracts with 32P-rs1271572G probe was inhibited by cold rs1271572G probes(×50 times higher than 32P labeled probes) (lane 15). The DNA-protein complex was supershifted after further incubating with anti-YY1 antibody in MDA-MB-231 cells (lane 16). Position A indicated the binding complex in the presence of the anti-YY1 antibody. Position B indicated the binding complex of 32Prs1271572G DNA probes with nuclear proteins from the different breast cell lines. Position C indicated free 32P- rs1271572G DNA Probes.

Mentions: To explore the role of the T allele in YY1 binding, binding activity of the two probes that contain sequence from either rs1271572G or rs1271572T was compared using EMSA in a panel of breast cancer cell lines. As shown in Figure 5, specific protein-DNA complex were detected with probe rs1271572G but not probe rs1271572T. Moreover, supershift with YY1 antibody indicated that YY1 was specifically bound to the rs1271572G oligo in these cells. Our data have shown that except for YY-1, other transcriptional factors could not bind to the region of rs1271572G and that YY-1 lost its capacity to bind to the region of rs1271572(T) (Figure 5).


Significance of rs1271572 in the estrogen receptor beta gene promoter and its correlation with breast cancer in a southwestern Chinese population.

Chen L, Liang Y, Qiu J, Zhang L, Chen X, Luo X, Jiang J - J. Biomed. Sci. (2013)

Binding of nuclear proteins in breast cancer cells to32 P- rs1271572G DNA probes. Different doses of nuclear proteins isolated from MDA-MB-231 cells were incubated with 32P-rs1271572G probes and the DNA-protein complexes were shown following autoradiography (lanes 1–7). The nuclear proteins isolated from MCF-7, MDA-MB-468, BT-549 and ZR75-30 were incubated with 32P -rs1271572G and the DNA-protein complexes were shown (lane 9–12). When 32Prs1271572T probes were used, no specific DNA-protein binding was observed (lanes 13 and 14). Binding activity of MDA-MB-231 nuclear extracts with 32P-rs1271572G probe was inhibited by cold rs1271572G probes(×50 times higher than 32P labeled probes) (lane 15). The DNA-protein complex was supershifted after further incubating with anti-YY1 antibody in MDA-MB-231 cells (lane 16). Position A indicated the binding complex in the presence of the anti-YY1 antibody. Position B indicated the binding complex of 32Prs1271572G DNA probes with nuclear proteins from the different breast cell lines. Position C indicated free 32P- rs1271572G DNA Probes.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3672062&req=5

Figure 5: Binding of nuclear proteins in breast cancer cells to32 P- rs1271572G DNA probes. Different doses of nuclear proteins isolated from MDA-MB-231 cells were incubated with 32P-rs1271572G probes and the DNA-protein complexes were shown following autoradiography (lanes 1–7). The nuclear proteins isolated from MCF-7, MDA-MB-468, BT-549 and ZR75-30 were incubated with 32P -rs1271572G and the DNA-protein complexes were shown (lane 9–12). When 32Prs1271572T probes were used, no specific DNA-protein binding was observed (lanes 13 and 14). Binding activity of MDA-MB-231 nuclear extracts with 32P-rs1271572G probe was inhibited by cold rs1271572G probes(×50 times higher than 32P labeled probes) (lane 15). The DNA-protein complex was supershifted after further incubating with anti-YY1 antibody in MDA-MB-231 cells (lane 16). Position A indicated the binding complex in the presence of the anti-YY1 antibody. Position B indicated the binding complex of 32Prs1271572G DNA probes with nuclear proteins from the different breast cell lines. Position C indicated free 32P- rs1271572G DNA Probes.
Mentions: To explore the role of the T allele in YY1 binding, binding activity of the two probes that contain sequence from either rs1271572G or rs1271572T was compared using EMSA in a panel of breast cancer cell lines. As shown in Figure 5, specific protein-DNA complex were detected with probe rs1271572G but not probe rs1271572T. Moreover, supershift with YY1 antibody indicated that YY1 was specifically bound to the rs1271572G oligo in these cells. Our data have shown that except for YY-1, other transcriptional factors could not bind to the region of rs1271572G and that YY-1 lost its capacity to bind to the region of rs1271572(T) (Figure 5).

Bottom Line: The rs1271572 G→T SNP abrogated YY1 binding and reduced the transcription activity of the promoter 0 N in the ERβ gene in vitro.TT genotype of rs1271572 inhibited expression of ERβ gene by down regulating transcriptional activity of the promoter 0 N in the ERβ gene.Our data revealed that the TT genotype of rs1271572 resulted in loss of the YY1 binding site and reduced the transcription activity of the promoter 0 N in the ERβ gene.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Background: To characterize single nucleotide polymorphisms (SNPs) within the promoter region of the estrogen receptor beta (ERβ) gene and to analyze the association of ERβ SNPs with susceptibility to breast cancer. Genotype frequencies of five SNPs (rs3020449, rs3020450, rs2987983, rs1271572 and rs1887994) in the promoter region of the ERβ gene in 873 women with breast cancer, 645 women with fibroadenoma and 700 healthy women were determined using an allele-specific tetra-primer polymerase chain reaction (PCR). Kaplan-Meier survival analysis was performed to evaluate the association of selected rs1271572 with prognosis in breast cancer. Electrophoretic mobility-shift assays were conducted to explore the binding of SNP rs1271572 containing probes to transcriptional factor Ying Yang 1 (YY1).

Results: Women with the homozygous TT genotype of rs1271572 had a significantly higher risk in developing breast cancer. Breast cancer patients with the TT genotype of rs1271572 had lower five-year survival rates than those with other genotypes and were more likely to suffer brain metastases. The rs1271572 G→T SNP abrogated YY1 binding and reduced the transcription activity of the promoter 0 N in the ERβ gene in vitro.

Conclusions: TT genotype of rs1271572 is associated with increased risk for breast cancer in Chinese women and is associated with unfavored prognosis in Chinese breast cancer patients. TT genotype of rs1271572 inhibited expression of ERβ gene by down regulating transcriptional activity of the promoter 0 N in the ERβ gene. Our data revealed that the TT genotype of rs1271572 resulted in loss of the YY1 binding site and reduced the transcription activity of the promoter 0 N in the ERβ gene.

Show MeSH
Related in: MedlinePlus