TINCR通过HuR和CeRNA双重作用上调EGFR表达促进乳腺癌细胞转移机制研究

Metastasis is the major cause of death in breast cancer patients. However, the mechanism remains unclear. Our previous studies showed that long non-coding RNA TINCR was upregualted significantly in more invasive breast cancer subtypes via high-thoughput sequencing (HTS) technology. Knockdown of TINCR deceased breast cancer cell migration and invasion in vitro and also suppressed lung metastasis in vivo. Next, HuR was identified through mass spectrometric analysis followed by RNA pull down assays. Furthermore, MIR708 and MIR125B2 were upregulated with knockdown of TINCR via HTS. Moreover, a CeRNA model is dectect between TINCR and EGFR via MIR708 and MIR125B2 examination. Multifunctional ASO/AS1411/Cy5.5 nanoparticles designed by us independently suppressed tumor effectively via targeting TINCR in vivo and MMTV-PyMT mice would be arranged to test the performance again. Thus, based on the results from previous study, we speculate that TINCR promotes EGFR expression via TINCR-HuR/MicroRNA-EGFR pathways. Integration analysis of TCGA、ICGC、GEO and ENCODE data, examination of breast cancer tissues in bio bank, mass spectrometric analysis, RIP, RNA pull down, Luciferase reporter assay, MMTV-PyMT mouse and multifunctional ASO/AS1411/Cy5.5 nanoparticle would be applied respectively to elucidate the potential molecular mechanism, enlightening a new orientation in exploring the function of TINCR, providing a new potential therapeutic target for breast cancer cell metastasis.

转移是乳腺癌病人死亡的主要原因。但是，机制并不完全清楚。申请人对各亚型乳腺癌样本行高通量转录组测序，识别lncRNA TINCR在高度转移性亚型中显著上调表达。敲降TINCR，乳腺癌细胞迁移与侵袭能力降低，肺转移瘤减少。通过测序及验证实验，明确TINCR靶基因EGFR及二者通过MIR708及MIR125B2构成CeRNA关系；RNA pull down后行质谱检测初步揭示TINCR与HuR结合。自主设计多功能纳米递送材料靶向TINCR效果佳。因此，提出TINCR-HuR/MicroRNA-EGFR轴促进EGFR表达科学假说。为证实该假说，项目将整合公共数据、临床样本、质谱、RIP、RNA pull down、Luciferase reporter、MMTV-PyMT转基因鼠、体内成像及纳米递送材料等实验手段，多层面、多角度系统研究，阐明TINCR促进乳腺癌细胞转移分子机制，提供治疗新靶点。

转移作为乳腺癌病人死亡的主要原因，其机制目前尚不完全清楚。申请人前期通过高通量转录组测序及相关验证实验、RNA pull down及质谱检测等实验结果，提出TINCR-HuR/MicroRNA-EGFR轴促进EGFR表达科学假说。为证实该假说，后续项目全面整合公共数据、临床样本、质谱、RIP、RNA pull down、Luciferase reporter、Balb/c小鼠体内荷瘤等实验手段，多层面、多角度系统研究，阐明了STAT3–TINCR–EGFR轴促进乳腺癌细胞转移的具体分子机制，为乳腺癌提供治疗新靶点。