L-NAME调控EMT抑制卵巢癌细胞迁移

目的探讨一氧化氮合酶泛抑制剂左旋硝基精氨酸甲基酯(L-NAME)对卵巢癌SKOV3细胞体外增殖、迁移以及对上皮间充质转化(EMT)标志物的影响。方法利用L-NAME抑制卵巢癌SKOV3中一氧化氮合酶(NOS)功能,以L-NAME处理组为实验组,二甲基亚砜(DMSO)组为对照组,采用CCK8增殖实验、平板克隆形成试验、划痕试验和Transwell小室迁移实验检测L-NAME对卵巢癌SKOV3细胞增殖和迁移功能的影响;并利用荧光定量PCR和Western blot检测卵巢癌细胞上皮细胞标志物E-cadherin和间充质细胞标志物N-cadherin、Vimentin和Fibronectin的表达,观察L-NAME对卵巢癌SKOV3细胞EMT标志物的调控作用。结果 CCK8增殖实验和平板克隆形成实验显示L-NAME抑制SKOV3细胞增殖功能(P0.05);划痕实验和Transwell小室实验显示L-NAME抑制SKOV3细胞迁移功能(P0.05);荧光定量PCR和Western blot结果显示L-NAME上调E-cadherin的表达,下调N-cadherin、Vimentin和Fibronectin的表达(P0.05)。结论 L-NAME抑制卵巢癌SKOV3细胞增殖、迁移功能,调节EMT标志物变化。

L-NAME regulating epithelial-mesenchymal transition suppressed ovarian cancer metastasis

Objective To investigate the effect of L-NAME on the proliferation, migration and epithelial-mesenchymal transition（EMT） of human ovarian cancer cell line SKOV3 in vitro. Methods SKOV3 cell line was treated with the inhibitor of NOS isoforms（L-NAME）, while DMSO was used as control. CCK8 assay, Colony formation assay, Scratch test and transwell chambers assay were employed to study the effect of L-NAME on SKOV3 proliferation and migration. Fluorescence quantitative PCR（qRT-PCR） and western blot were employed to detect the changes of EMT related genes expression. Results Compared to control group CCK8 and Colony formation assay showed the proliferation of SKOV3 was significantly reduced after treated with L-NAME. Scratch test and transwell chambers migration experiment showed that L-NAME suppressed ovarian cancer cell migration. qRT-PCR and western blot showed that L-NAME up-regulated the expression of E-cadherin in SKOV3 cells, while down-regulated the expression of N-cadherin, Vimentin and Fibronectin. Conclusion L-NAME significantly inhibited proliferation, and migration of ovarian cancer SKOV3 cells in vitro, and suppressed EMT-like cellular marker alterations.