卵巢癌死亡率高居妇科恶性肿瘤之首，如何抑制卵巢癌细胞增殖及侵袭是提高其预后的重要途径之一。微小RNA（microRNA）作为一种转录后调节因子，参与细胞增殖和肿瘤侵袭，并早期释放入血。我们前期研究发现miR-24在卵巢癌患者表达改变，并可调节卵巢癌细胞的周期，癌组织内侵袭相关因子磷酸化蛋白激酶4（PAK4）和LIMK-1表达亦增加。结构生物信息学提示miR-24可作用于细胞骨架调节蛋白PAK4，继而调控LIMK1/cofilin通路而抑制细胞迁移来降低卵巢癌细胞增殖侵袭潜能。为此本研究拟检测不同临床分期及治疗方案的卵巢癌患者miR-24表达并随访，研究其对预后的诊断价值，同时建立miR-24上调和下调表达的卵巢癌细胞株和裸鼠动物模型，通过体内外实验研究不同miR-24水平对卵巢癌细胞增殖、侵袭以及调控PAK4/LIMK-1分子信号通路的作用，以期为卵巢癌治疗及预后预测提供新的思路与方法。

The mortality rates of ovarian cancer was the highest in the gynecological malignancies, and studies have shown that inhibiting ovarian cancer cell invasion and metastasis is an important way to improve the prognosis. MicroRNA as a transcriptional regulatory factor involved in cell proliferation and tumor invasion, and early release into the blood. Our previous study found that the expression of miR-24 was changed in the ovarian cancer tissue. miR-24 could adjust the cycle of ovarian cancer cells. The phosphorylation of protein kinase 4 (PAK4) and LIMK-1 expression in ovarian cancer tissue were also changed. The structural biology suggested miR-24 could regulate and contral expression of the PAK4 mRNA, then inhibiting LIMK1/cofilin pathway and eventually reducing the potential ability of ovarian cancer cell proliferation and migration. Therefore, this study will detect the miR-24 expresion in the different clinical stage and treatment options in patients with ovarian cancer, and follow-up the cases to study its predictive value as a prognoses. We will up and down-regulate the expression of miR-24 respectively by using sence sequence and antisense sequence RNA oligonucleotide to construct lentiviral expression vector,which were transfected into the epithelial ovarian cancer cell lines, and induce in vitro severe combined immunodeficiency mice xenograft tumor model. The present study will explore the miR-24 targeting in adjustment the above-mentioned signaling pathways and evaluate effects on cell proliferation and invasion. By the in vitro and in vivo experiments, we could confirm these bioinformatics prediction above mentioned, and inhibit ovarian cancer cell proliferateive and invasive potential from the gene level ,thereby, to provide the new way for the treatment and predict prognosis of ovarian cancer.

上皮性卵巢癌作为致死率最高的妇科肿瘤，晚期患者的5年生存率不到30%。lncRNA和 miRNA是两类常见的非编码RNA，参与转录调控、细胞增殖、分化等多种生物学过程，在肿瘤相关的基因调控系统中扮演着关键的角色。在本研究中，我们首先利用基因芯片检测发现LINC01088在良性上皮性卵巢肿瘤中的表达较正常卵巢上皮组织中明显减少，并且在42个临床样本中进一步得到了验证，且在上皮性交界性和恶性卵巢肿瘤中的表达量下降更加明显。为了确认LINC01088在上皮性卵巢肿瘤发生中的角色，我们构建过表达LINC01088的慢病毒载体并建立高表达LINC01088细胞系，进一步的动物致瘤实验表明表达LINC01088能抑制动物移植瘤的生长。接着我们通过临床样本相关性分析和RNA共沉淀方法验证了miR-24-1-5p是LINC01088作用的靶点，提示LINC01088可能是通过miR-24-1-5p而抑制细胞生长的。接着，我们建立了稳定表达miR-24-1-5p的A2780细胞系进行了细胞增殖实验、细胞迁移实验以及皮下移植瘤实验，结果表明表达miR-24-1-5p在体内外均能促进细胞增殖，而LINC01088能削弱表达miR-24-1-5p对细胞的促增殖作用，但表达miR-24-1-5p并不促进细胞的迁移。最后我们还通过报告基因和WB方法验证了PAK4是miR-24-1-5p下游靶蛋白，且细胞增殖实验提示表达PAK4显著抑制细胞的增殖，而细胞迁移实验表明过表达PAK4能增强细胞迁移能力。通过本研究培养硕士研究生4名，申请专利1项（审批中），已发表SCI论文2篇，已发表国内核心期刊论文1篇，已收录SCI论文1篇，在投SCI论文2篇（1篇已修回）。