VEGF111b对卵巢癌生长和血管化的作用与相关机制研究

The high mortality and recurrence rate of ovarian cancer is a problem in clinical treatment,and angiogenesis plays an important role. After VEGF gene is differentially spliced, it can result in proangiogenic VEGFxxx isoforms and antiangiogenic VEGFxxxb isoforms. This study has confirmed that the mRNA of VEGF111b, a new VEGF splicing isoform, exists in the SKOV3 cells and ovarian cancer tissues. We have found there is differenent expression of VEGF111b between the normal ovarian tissues and ovarian cancer tissues.We will prepare recombinant protein VEGF111b and monoclonal antibody VEGF111b, and detect the expression of VEGF111b in ovarian cancer tissues and blood serum. Experiments in vitro will be used to test whether recombinant protein VEGF111b can inhibit tumor cell growth and angiogenesis.Through the above study, preliminarily we want to know the effects of VEGF111b in inhibiting growth and angiogenesis on ovarian cancer, so as to provide a high-efficiency angiogenesis inhibitors for clinical treatment of ovarian cancer.

卵巢癌高死亡率和复发率是临床治疗面临的难题，血管生成在其中发挥重要作用。VEGF基因差异剪接后能够产生促进血管生成的VEGFxxx和抑制血管生成的VEGFxxxb。本研究已证实一种新型的剪接差异变异体VEGF111b的mRNA在SKOV3细胞和卵巢癌组织中存在，并在正常卵巢和卵巢癌组织中存在差异性表达；在此基础上，制备VEGF111b单克隆抗体及ELASA试剂盒，在蛋白水平上检测其在卵巢癌组织及血清中的表达；制备VEGF111b、VEGF165b及VEGF121b重组融合蛋白，通过体外实验研究重组VEGF111b抑制肿瘤生长及血管化的作用，且活性比VEGF165b、VEGF121b强。通过上述研究，初步了解VEGF111b抑制卵巢癌生长和血管化的作用，以期为临床卵巢癌治疗提供一种高效低毒的抗血管生成抑制剂。

我们通过使用VEGF111b特异性引物进行RT-PCR证实VEGF111b在卵巢癌细胞中诱导性表达，制备了真核表达载体pcDNA3.1-VEGF111b和多克隆抗体，并通过western blot证明VEGF111b蛋白在卵巢癌细胞中也是诱导性表达。通过将VEGF111b过表达，我们发现VEGF111b过表达后能够抑制人脐带内皮细胞HUVEC的增殖、迁移及小管形成，并初步探讨了VEGF111b体外抑制血管形成的机制，主要是通过VEGF-R2/PI3K/Akt 和VEGF-R2/ERK1/2信号通路的抑制。同时，发现现VEGF111b过表达后也能够抑制卵巢癌SKOV3细胞的增殖及克隆形成，并发现使SKOV3细胞周期阻滞在S期。