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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(P<0.05);划痕实验和Transwell小室实验显示L-NAME抑制SKOV3细胞迁移功能(P<0.05);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(P<0.05)。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。
Objective To investigate the effects of L-NAME, a nitric oxide synthase inhibitor, on the proliferation, migration and epithelial mesenchymal transition (EMT) markers in ovarian cancer SKOV3 cells in vitro. Methods L-NAME was used to inhibit nitric oxide synthase (NOS) function in ovarian cancer cell line SKOV3. L-NAME group and DMSO group were used as control group. CCK8 proliferation assay, Formation assay, scratch assay and Transwell chamber migration assay were used to detect the effect of L-NAME on the proliferation and migration of ovarian cancer SKOV3 cells. Fluorescent quantitative PCR and Western blot were used to detect the expression of E-cadherin and mesenchyme Cell markers N-cadherin, Vimentin and Fibronectin, to observe the effect of L-NAME on EMT markers in ovarian cancer SKOV3 cells. Results Both L-NAME and L-NAME inhibited the proliferation of SKOV3 cells (P <0.05). Scratch assay and Transwell chamber assay showed that L-NAME inhibited the migration of SKOV3 cells (P <0.05) Western blot results showed that L-NAME up-regulated the expression of E-cadherin and down-regulated the expression of N-cadherin, Vimentin and Fibronectin (P <0.05). Conclusion L-NAME can inhibit the proliferation and migration of ovarian cancer SKOV3 cells and regulate the changes of EMT markers.