临床与病理杂志2017，Vol.37Issue(7)：1331-1340,10.DOI:10.3978/j.issn.2095-6959.2017.07.002
ORP-150基因沉默对SKOV-3细胞增殖、迁移及侵袭的影响
Effect of ORP-150 gene silencing on proliferation, migration and invasion of SKOV-3 cells
摘要
Abstract
Objective: To investigate the effect of silencing ORP-150 gene on the proliferation and metastasis of ovarian cancer cell line SKOV-3 in vitro and to explore its possible mechanism.Methods: The ORP-150 gene of SKOV-3 cells was silenced by lentivirus packaging shRNA method, and then 3 groups were designed: An experimental group (ORP-150 shRNA), a negative control group (NC shRNA) and a blank control group (CON). qPCR and Western Blot were used to detect gene knockout efficiency. CCK-8 method and clone formation assay were used to detect the effect of ORP-150 silencing on cell proliferation ability. Wound Healing and transwell tests were used to detect the invasion and metastasis of ovarian cancer cells. Flow cytometry was used to detect the apoptosis of cells. Western-blot was used to investigate the possible mechanism of ORP-150 gene knockdown on cell phenotype changes.Results: Silencing ORP-150 gene could inhibit the proliferation ability, clone formation ability and transmembrane transfer and diflusion ability of ovarian cancer cells (P<0.05), but had no signiflcant effect on the invasive ability of the cells (P>0.05). The apoptosis of ovarian cancer cells was significantly increased afler knockdown of ORP-150 gene (P<0.05). Ovarian cancer cells knocked down ORP-150 gene may aflect the cell repair through Wnt signalling pathway and Caspase cascade reaction, and ultimately inhibit cell proliferation and promote apoptosis.Conclusion: Silencing ORP-150 gene inhibits the proliferation and metastasis of ovarian cancer cell line SKOV-3 and promotes cell apoptosis. This effect may be achieved by inhibiting the expression of c-myc, cyclin D1 and caspase-3 proteins through inhibiting the expression of β-catenin in the Wnt signalling pathway and inhibiting the Caspase cascade. ORP-150 gene silencing may be a new target for ovarian cancer treatment.关键词
上皮性卵巢癌/ORP-150/细胞增殖/Wnt信号通路/凋亡Key words
epithelial ovarian cancer/ORP-150/cell proliferation/Wnt signalling pathway/apoptosis引用本文复制引用
孙中峰,张萍..ORP-150基因沉默对SKOV-3细胞增殖、迁移及侵袭的影响[J].临床与病理杂志,2017,37(7):1331-1340,10.基金项目
上海市科学技术委员会项目(14ZR1426700). This work was supported by Foundation of Shanghai Science and Technology Grant, China (14ZR1426700). （14ZR1426700）
