前列腺癌骨转移低表达miRNA调控KLF17促进骨转移的机制研究

The most common site of PCa metastasis is the bone, with bone metastases identified at autopsy in up to 90%. The mechanism of miRNA modulate EMT and stemness in bone metastasis is unclear. Low-expressed of miR-100,-145,-143 and high-expressed of KLF17 in PCa are related with bone metastasis of patient, and can promote tumor cell EMT and stemness, respectively. migration to the tumor site, and subsequent integration into the vascular network. As a result, we speculate in prostate cancer bone metastases down-regulation miRNA promoted KLF17 expression ,modulated EMT and stemness,and induced bone metastasis.So, in this program we intend to high expressed miRNA, and transfected siRNA and high expression KLF17 plasmid to affect KLF17 expression, and identify the influence on cell form, functions of proliferation, sphere, colony-formation,adhesion,migration.invasive, the mark expression of EMT and stmness in vitro; Meanwile, testify miRNA and KLF17 influence on bone metatasis in vivo by tibial injection and heart injection module of nude mice, through dectect bone destruction by X-ray and MR, on mark expression of EMT and stemness by HE and immunohistochemistry .Eventually, we want to clarify the bone metastasis mechanism of miRNA regulated the KLF17 in PCa, thus providing important basis for studying the prevention and therapy of PCa bone metasis.

骨转移是前列腺癌(PCa)患者主要死因。miRNA在骨转移中调控肿瘤内皮间充质化（EMT）和干性，但机制仍不清楚。前期临床样本发现PCa低表达miR-100，-145，-143，高表达其靶基因KLF17,均与骨转移密切相关，且均能促进骨转移细胞EMT和干性。据此假设上述下调的miRNA通过上调靶基因KLF17，调控EMT和干性，促进骨转移。本项目通过在骨转移PC-3细胞中高表达这些miRNA，和转染siRNA或高表达质粒影响KLF17的表达，观察其对骨转移细胞体外形态、增殖、成球、集落形成、粘附、迁移、侵袭、EMT和干性标志物表达的影响；体内通过小鼠胫骨和心脏注射模型验证miRNA及靶基因KLF17对骨转移的影响，X光、MR检测骨破坏，HE染色和免疫组化检测骨转移瘤EMT和干性标志物表达。阐明骨转移低表达miRNA调控KLF17促进骨转移的机制，为探寻PCa骨转移预防和治疗的新靶标提供依据

骨转移是前列腺癌(PCa)患者主要死因。miRNA在骨转移中调控肿瘤内皮间充质化（EMT）和干性，但机制仍不清楚。前期临床样本发现PCa低表达miR-100，-145，-143，高表达其靶基因KLF17,均与骨转移密切相关，且均能促进骨转移细胞EMT和干性。据此假设上述下调的miRNA通过上调靶基因KLF17，调控EMT和干性，促进骨转移。本项目通过在骨转移PC-3细胞中高表达这些miRNA，和转染siRNA或高表达质粒影响KLF17的表达，观察其对骨转移细胞体外细胞功能学的影响；体内通过小鼠心脏注射模型验证miRNA及靶基因KLF17对骨转移的影响，X光、HE染色检测骨破坏。结果表明，KLF17是miR-145，-143的直接靶基因，miR-145，-143的低表达能够影响PCa骨转移细胞的形态，促进细胞增殖，体外迁移和侵袭，同时也能影响PCa骨转移细胞EMT和干性标志物表达。进一步的动物实验证明，anti-miRNA，siKLF17对骨转移的作用，X光、MR检测了骨破坏，HE染色和免疫组化检测了骨转移。阐明骨转移低表达miRNA调控KLF17促进骨转移的机制，为探寻PCa骨转移预防和治疗的新靶标提供依据.