CircFUT8竞争性结合miR-361-3p调控Paxillin介导胰腺癌吉西他滨耐药的机制研究

As a new research frontier of long noncoding RNAs, the mechanism of circular circRNAs in the regulation of pancreatic cancer drug resistance is still unclear. We previously found that expression of circFUT8 was significantly increased in gemcitabine-resistant pancreatic cancer cells, whereas RNA interference of circFUT8 reversed the drug resistance. The majority of circFUT8 was present in the cytoplasm, and miR-361-3p may directly bind to circFUT8. Besides, Paxillin (PXN) was significantly decreased after transfection of circFUT8 small interfering RNA or miRNA-361-3p mimic. Based on these findings, we hypothesize that high expression of circFUT8 plays an important role in the acquired gemcitabine-resistance of pancreatic cancer, which function as a sponge for miRNA-361-3p and abrogated their suppression of PXN. To validate this hypothesis, we will determine the role of circFUT8 and study the mechanism in acquired gemcitabine-resistance of pancreatic cancer with multidisciplinary approaches of molecular, cell biology and bio-information in terms of cell level, animal model and clinical level. The project will provide new therapeutic targets and biomarker to overcome the gemcitabine-resistance of pancreatic cancer.

CircRNAs做为非编码RNA研究的前沿领域和热点，其在胰腺癌耐药中的分子机制尚不清楚。我们前期研究发现circFUT8在胰腺癌吉西他滨耐药细胞中表达上调，干扰circFUT8的表达能逆转吉西他滨耐药；circFUT8主要分布于细胞质中且能与miR-361-3p结合；细胞水平干扰circFUT8或转染miRNA-361-3p mimic均能下调PXN表达。据此我们提出假说：circFUT8的表达上调可能是诱导胰腺癌获得性耐药的重要原因，它可能作为miRNA-361-3p sponge减弱miRNA-361-3p对PXN的抑制作用介导耐药。为验证这一假说，本项目拟从细胞、动物和临床水平，综合分子、细胞生物学、生物信息学等学科交叉手段，研究circFUT8的差异表达对耐药的影响，探讨其介导胰腺癌吉西他滨耐药的分子机制，为逆转胰腺癌吉西他滨耐药提供新的靶点和分子标志物。

频发的耐药现象已成为胰腺癌吉西他滨（GEM）治疗的瓶颈问题，耐药患者肿瘤非编码RNA呈现明显的差异表达，提示非编码RNA在肿瘤耐药中可能发挥重要的作用，然其作用机制还有待深入的研究。本课题利用构建的胰腺癌GEM耐药细胞株，通过RNA-Seq筛选并验证了耐药细胞中差异表达的circRNA及miRNA，找到四个GEM耐药相关的circRNA和三个miRNA；发现circFUT8在胰腺癌GEM耐药细胞表达显著上调，干扰circFUT8表达能逆转GEM耐药，抑制耐药细胞增殖、迁移；另外miR-3065-5p可以增加胰腺癌耐药细胞对GEM的敏感性，耐药细胞中下调的miR-3065-5p可能通过减弱其对HMOX1的抑制作用进而诱导GEM耐药。本研究对胰腺癌GEM耐药相关差异表达的circRNA和miRNA做了深入的分析和研究，找到了几种胰腺癌GEM耐药过程中发挥重要作用的非编码RNA，不仅为逆转GEM耐药提供新的理论依据和治疗策略，还为解决胰腺癌耐药难题提供新的靶点和分子标志物。

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