Neoplasma. 2016;63(5):696-704. doi: 10.4149/neo_2016_506.
We aimed to determine the effects of miR-433 on the malignant behaviors of ovarian cancer cells, as well as to elucidate the possible mechanisms of ovarian cancer development. A total of 9 ovarian cancer tissues and 9 matched normal ovary tissues were obtained, and the expression levels of miR-433 and Notch1 were then determined by real-time PCR. Human ovarian cancer cell lines SKOV3 and OVCAR3 were transfected with miR-433 mimics, negative miR-control and Notch1 siRNA. The expression of Notch1 protein in transfected cells was determined by western blot. In addition, the proliferation, migration and invasion of SKOV3 and OVCAR3 cells in vitro were then evaluated using Cell Counting Kit 8, wound healing assay and Transwell invasion assay, respectively. Besides, bioinformatics methods and luciferace reporter assay were performed to confirm whether Notch1 was a direct target of miR-433. The expression of miR-433 was markedly down-regulated while Notch1 expression was significantly up-regulated in ovarian cancer tissues compared with matched normal ovary tissues. Overexpression of miR-433 significantly inhibited the migration and invasion of ovarian cancer cells, but had not significant effects on cell proliferation. In addition, Notch1 was a direct target of miR-433. Besides, down-regulation of Notch1 inhibited the invasion of ovarian cancer cells. Our findings indicate that miR-433 may inhibit cell migration and invasion in the development of ovarian cancer via down-regulation of Notch1. Notch1 may serve as a potential target in cancer therapy.
我们旨在确定 miR-433 对卵巢癌细胞恶性行为的影响，并阐明卵巢癌发展的可能机制。共获得 9 例卵巢癌组织和 9 例配对的正常卵巢组织，然后通过实时 PCR 测定 miR-433 和 Notch1 的表达水平。用 miR-433 模拟物、阴性 miR 对照和 Notch1 siRNA 转染人卵巢癌细胞系 SKOV3 和 OVCAR3。通过 Western blot 测定转染细胞中 Notch1 蛋白的表达。此外，分别通过细胞计数试剂盒 8、划痕愈合试验和 Transwell 侵袭试验评估 SKOV3 和 OVCAR3 细胞在体外的增殖、迁移和侵袭。此外，通过生物信息学方法和荧光素酶报告基因检测证实 Notch1 是否是 miR-433 的直接靶标。与配对的正常卵巢组织相比，卵巢癌组织中 miR-433 的表达明显下调，而 Notch1 的表达明显上调。miR-433 的过表达显著抑制卵巢癌细胞的迁移和侵袭，但对细胞增殖没有显著影响。此外，Notch1 是 miR-433 的直接靶标。下调 Notch1 抑制卵巢癌细胞的侵袭。我们的研究结果表明，miR-433 可能通过下调 Notch1 抑制卵巢癌的发展过程中的细胞迁移和侵袭。Notch1 可能是癌症治疗的潜在靶点。
