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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.

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The T allele of rs1271572 down-regulated the activity of the ERβ promoter 0 N.A). The human breast cancer cells (MDA-MB-231, MCF-7, MDA-MB-468, BT-549 and ZR75-30) were transfected with the normalization control vector pRL and each of the pESR2-0 N-G-Luc, pESR2-0 N-T-Luc, pESR2-0 K-Luc (promoter 0 K) alone, or together with the YY1-specific siRNA oligos. Lucifease activities of these reporters were determined using the dual luciferase assay kit. B). Primary breast cancer cells (N1, N2 and N3) were transfected with the vectors as in A) and luciferase activity was determined. The figure represents the results of three independent experiments. *p <0.05;**p <0.01. Graphs show means + −S.E.M. (n =8). C). The human breast cancer cell lines (MDA-MB-231, MCF-7, MDAMB468, BT549 and ZR75-30) and the primary breast cancer cells (N1 and N2) were transfected with YY1-specific siRNA oligos and YY1 expression was examined by western blot. β-actin was used to confirm equal protein loading. Each lane was loaded with up to 30 μg of protein.
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Figure 6: The T allele of rs1271572 down-regulated the activity of the ERβ promoter 0 N.A). The human breast cancer cells (MDA-MB-231, MCF-7, MDA-MB-468, BT-549 and ZR75-30) were transfected with the normalization control vector pRL and each of the pESR2-0 N-G-Luc, pESR2-0 N-T-Luc, pESR2-0 K-Luc (promoter 0 K) alone, or together with the YY1-specific siRNA oligos. Lucifease activities of these reporters were determined using the dual luciferase assay kit. B). Primary breast cancer cells (N1, N2 and N3) were transfected with the vectors as in A) and luciferase activity was determined. The figure represents the results of three independent experiments. *p <0.05;**p <0.01. Graphs show means + −S.E.M. (n =8). C). The human breast cancer cell lines (MDA-MB-231, MCF-7, MDAMB468, BT549 and ZR75-30) and the primary breast cancer cells (N1 and N2) were transfected with YY1-specific siRNA oligos and YY1 expression was examined by western blot. β-actin was used to confirm equal protein loading. Each lane was loaded with up to 30 μg of protein.

Mentions: To further determine whether the T allele of rs1271572 confers decreased transcriptional activity of the ERβ gene promoter, we compared the luciferase reporter activity of pESR2-0 N-G-Luc and pESR2-0 N-T-Luc. Indeed, the reporter activity of pESR2-0 N-T-Luc was significantly lower that of pESR2-0 N-G-Luc, (Figure 6A and6B), suggesting that the T allele of rs1271572 could lead to a decrease in the transcription activity of the ERβ gene promoter 0 N. In agreement with a previous report that transcripts from promoter 0 N were more prominent than those from promoter 0 K[16], we also observed that the luciferase activity of pESR2-0 K-Luc was significantly weaker than that of pESR2-0 N-G-Luc in the five breast cancer cell lines and in the primary cancer cells of two patients. Interestingly, luciferase activity of pESR2-0 N-G-Luc but not pESR2-0 N-T-Luc was dramatically decreased in these cells when co-transfected with YY1-specific siRNA oligos along with pESR2-0 N-G-Luc, pESR2-0 N-T-Luc or pESR2-0 K-Luc, indicating that YY1 is involved in regulation of the activity of ERβ gene promoter 0 N (Figure 6A and6B).


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)

The T allele of rs1271572 down-regulated the activity of the ERβ promoter 0 N.A). The human breast cancer cells (MDA-MB-231, MCF-7, MDA-MB-468, BT-549 and ZR75-30) were transfected with the normalization control vector pRL and each of the pESR2-0 N-G-Luc, pESR2-0 N-T-Luc, pESR2-0 K-Luc (promoter 0 K) alone, or together with the YY1-specific siRNA oligos. Lucifease activities of these reporters were determined using the dual luciferase assay kit. B). Primary breast cancer cells (N1, N2 and N3) were transfected with the vectors as in A) and luciferase activity was determined. The figure represents the results of three independent experiments. *p <0.05;**p <0.01. Graphs show means + −S.E.M. (n =8). C). The human breast cancer cell lines (MDA-MB-231, MCF-7, MDAMB468, BT549 and ZR75-30) and the primary breast cancer cells (N1 and N2) were transfected with YY1-specific siRNA oligos and YY1 expression was examined by western blot. β-actin was used to confirm equal protein loading. Each lane was loaded with up to 30 μg of protein.
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Figure 6: The T allele of rs1271572 down-regulated the activity of the ERβ promoter 0 N.A). The human breast cancer cells (MDA-MB-231, MCF-7, MDA-MB-468, BT-549 and ZR75-30) were transfected with the normalization control vector pRL and each of the pESR2-0 N-G-Luc, pESR2-0 N-T-Luc, pESR2-0 K-Luc (promoter 0 K) alone, or together with the YY1-specific siRNA oligos. Lucifease activities of these reporters were determined using the dual luciferase assay kit. B). Primary breast cancer cells (N1, N2 and N3) were transfected with the vectors as in A) and luciferase activity was determined. The figure represents the results of three independent experiments. *p <0.05;**p <0.01. Graphs show means + −S.E.M. (n =8). C). The human breast cancer cell lines (MDA-MB-231, MCF-7, MDAMB468, BT549 and ZR75-30) and the primary breast cancer cells (N1 and N2) were transfected with YY1-specific siRNA oligos and YY1 expression was examined by western blot. β-actin was used to confirm equal protein loading. Each lane was loaded with up to 30 μg of protein.
Mentions: To further determine whether the T allele of rs1271572 confers decreased transcriptional activity of the ERβ gene promoter, we compared the luciferase reporter activity of pESR2-0 N-G-Luc and pESR2-0 N-T-Luc. Indeed, the reporter activity of pESR2-0 N-T-Luc was significantly lower that of pESR2-0 N-G-Luc, (Figure 6A and6B), suggesting that the T allele of rs1271572 could lead to a decrease in the transcription activity of the ERβ gene promoter 0 N. In agreement with a previous report that transcripts from promoter 0 N were more prominent than those from promoter 0 K[16], we also observed that the luciferase activity of pESR2-0 K-Luc was significantly weaker than that of pESR2-0 N-G-Luc in the five breast cancer cell lines and in the primary cancer cells of two patients. Interestingly, luciferase activity of pESR2-0 N-G-Luc but not pESR2-0 N-T-Luc was dramatically decreased in these cells when co-transfected with YY1-specific siRNA oligos along with pESR2-0 N-G-Luc, pESR2-0 N-T-Luc or pESR2-0 K-Luc, indicating that YY1 is involved in regulation of the activity of ERβ gene promoter 0 N (Figure 6A and6B).

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