Effect of functional variant rs11466313 on breast cancer susceptibility and TGFB1 promoter activity
Effect of functional variant rs11466313 on breast cancer susceptibility and TGFB1 promoter activity
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功能性变异rs11466313对乳腺癌易感性和TGFB1启动子活性的影响
DOI：
10.1007/s10549-020-05841-w
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发表时间：
2020-08-05
影响因子：
3.8
通讯作者：
Fang，Wei-Gang
中科院分区：
文献类型：
--
作者：
Zhou，Yan-Ting;Zheng，Li-Yuan;Fang，Wei-Gang
目的探讨TGF β 1基因多态性与乳腺癌易感性的关系，并探讨其作用机制。生物信息学分析用于预测与tSNPs紧密相关的功能SNP。进行荧光素酶基因报告基因测定以确定遗传变体对启动子活性的影响。结果TGFB 1基因5′端调控区rs 1800469（C>T）与BC风险降低相关，TGFB 1基因5′端调控区rs 1800469（C>T）与BC风险降低相关。生物信息学分析预测TGFB 1基因5′调控区rs 11466313（-2389_-2391 Del/AGG）与tSNP rs 1800469紧密连锁，可能具有功能。通过PCR-SSCP对rs 11466313进行基因分型，结果显示rs 11466313也可降低BC风险。荧光素酶分析表明，与其主要等位基因相比，rs 11466313次要等位基因的启动子活性降低超过9倍（p< 0.001）。DNA pull-down分析和质谱分析表明rs 11466313次要等位基因失去了与FAM 98 B和HSP 90 B的结合能力。敲低FAM 98 B，但不是热休克蛋白90 B，增强启动子活性驱动的TGFB 1 rs 11466313主要等位基因是attenued.ConclusionsThis研究阐明了影响的功能多态性rs 11466313在调节区TGFB 1乳腺癌的易感性和基因表达，并可能有助于未来的研究，以确定这种TGFB 1变异在临床环境中的价值。
PurposeThis study aimed to investigate whether genetic polymorphisms in TGFB1 contribute to breast cancer (BC) susceptibility, and explore the mechanism of action.MethodsA total of 7 tagging SNPs (tSNPs) were genotyped in 1161 BC cases and 1337 age-matched controls among Chinese Han population. Bioinformatics analysis was used to predict functional SNP closely linked to tSNPs. Luciferase gene reporter assay was performed to determine the effect of genetic variants on promoter activity. DNA pull-down assay and mass spectrometry were used to identify the differentially binding proteins to genetic variants.ResultsGenotyping analysis showed that rs1800469 (C>T) in the 5′ regulatory region of TGFB1 was associated with reduced BC risk. Bioinformatics analysis predicted that rs11466313 (-2389_-2391 Del/AGG) in the 5′ regulatory region of TGFB1, was closely linked to tSNP rs1800469 and could be functional. The genotyping of rs11466313 by PCR-SSCP showed that rs11466313 also conferred decreased BC risk. Luciferase assays demonstrated that rs11466313 minor allele reduced over ninefold of promoter activity compared with its major allele (p< 0.001). DNA pull-down assay and mass spectrometry revealed that rs11466313 minor allele lost the binding ability with FAM98B and HSP90B. Knocking down FAM98B but not HSP90B, the enhanced promoter activity driven by TGFB1 rs11466313 major allele was attenuated.ConclusionsThis study elucidates the impact of functional polymorphism rs11466313 in the regulatory region of TGFB1 on breast cancer susceptibility and gene expression, and could be helpful for future research to determine the value of this TGFB1 variant in the clinical setting.