乳腺癌细胞中HuR调控PDFC的分子机制研究

目的:探讨乳腺癌细胞中HuR调控PDGFC的分子机制。方法:通过软件预测分析,乳腺癌细胞中PDGFC 3'UTR的HuR结合位点;在RNA免疫共沉淀实验中,加入PDGFC刺激后检测HuR与PDGFC mRNA的相互作用;通过构建PDGFC 3'UTR五个截短体,荧光素酶报告基因实验检测HuR调控PDGFC的结合位点。结果:软件预测分析发现,PDGFC 3'UTR可能存在五个HuR结合位点;RNA免疫共沉淀实验中,当加入PDGFC刺激后,HuR与PDGFC mRNA出现免疫共沉淀,证明HuR和PDGFC之间的直接相互作用;PDGFC mRNA3'UTR报告基因系统检测显示,第2个和第4个位点可与HuR结合调控PDGFC。结论:本研究揭示了乳腺癌细胞中HuR通过与PDGFC mRNA 3'UTR结合调控PDGFC的分子机制,为乳腺癌的临床诊断和治疗提供了新的思路。

Molecular Mechanism of PDGFC Regulated by HuR in Breast Cancer Cells

ABSTRACT Objective: To investigate the molecular mechanism of PDGFC regulated by HuR in breast cancer cells. Methods: Through the software analysises, we first predicted the HuR-binding sites on the PDGFC 3''-UTR in breast cancer cells; An RNA-im- munoprecipitation tested the interaction of HuR with the PDGFC mRN after adding PDGFC stimulation; Luciferase experiments tested the HuR-binding sites of PDGFC regulated by HuR through structuring five truncated of PDGFC 3''UTR. Results: We found five HuR-binding sites in the 3''UTR of PDGFC by software prediction; The RNA-immunoprecipitation showed the co-immunoprecipitation of HuR and PDGFC mRN after adding PDGFC stimulation confirming the direct association of HuR with the PDGFC; Luciferase experi- ments of PDGFC mRNA 3''UTR showed that PDGFC regulated by the second and fourth HuR-binding sites. Conclusion: This study re- veals the molecular mechanism of PDGFC regulated by HuR through binding to PDGFC mRNA 3''UTR in breast cancer cells, and pro- vided rationale for the development of strategies in the clinical diagnosis and treatment for breast cancer.