ABSTRACT
@#[摘 要] 目的：探讨吴茱萸碱（Evo）是否通过调控lncRNA LINC00858表达调控神经母细胞瘤SK-N-SH细胞的增殖、迁移及侵袭。方法：在体外以3、6、12 μmol/L Evo处理人神经母细胞瘤SK-N-SH细胞，利用RNA干扰技术分别将si-NC、si-LINC00858转染至SK-N-SH细胞，将pcDNA、pcDNA-LINC00858转染至SK-N-SH细胞并经12 μmol/L Evo处理，实验分为对照组、Evo低剂量组、Evo中剂量组、Evo高剂量组、si-NC组、si-LINC00858组、Evo+pcDNA组、Evo+pcDNA-LINC00858组。采用qPCR法检测各组细胞LINC00858的表达量，MTT、Transwell实验分别检测细胞的增殖、迁移、侵袭能力，WB法检测细胞中cyclinD1、MMP-2、MMP-9和p21蛋白的表达。结果：与对照组相比，Evo低、中、高剂量组SK-N-SH细胞中LINC00858表达均显著降低（均P<0.05），细胞增殖抑制率显著升高、迁移及侵袭细胞数显著减少（均P<0.01），cyclinD1、MMP-2、MMP-9蛋白表达降低、p21蛋白表达升高（均P<0.01）。与si-NC组相比，si-LINC00858组细胞的增殖抑制率、迁移和侵袭细胞数及相关蛋白表达变化同Evo低、中、高剂量组。与Evo+pcDNA组相比，Evo+pcDNA-LINC00858组细胞的增殖抑制率显著降低、迁移及侵袭细胞数均显著增多（均P<0.01），cyclinD1、MMP-2、MMP-9蛋白表达升高、p21蛋白表达降低（均P<0.05）。结论：Evo通过下调LINC00858表达抑制神经母细胞瘤SK-N-SH细胞的增殖、迁移及侵袭。
ABSTRACT
Antitumor effects of erythromycin and the related mechanism were investigated in the present study.Neuroblastoma cells (SH-SY5Y) were exposed to erythromycin at different concentrations for different durations.Cell proliferation was measured by cell counting,and cell viability was examined by MTT assay.Cell cycle phase distribution and the cytosolic calcium level were detected by flow cytometry.Mitochondrial membrane potential was measured by the JC-1 probe staining and fluorescent microscopy.The expression of an oncogene (c-Myc) and a tumor suppressor [p21 (WAF1/Cipl)] proteins was analyzed by using Western blotting.Erythromycin could inhibit the proliferation of SH-SY5Y cells in a concentration- and time-dependent manner.The cell cycle was arrested at S phase.Mitochon drial membrane potential collapsed and the cytosolic calcium was overloaded in SH-SY5Y cells when treated with erythromycin.The expression of c-Myc protein was down-regulated,while that of p21 (WAF1/Cip1) protein was up-regulated.It was concluded that erythromycin could restrain the proliferation of SH-SY5Y cells.The antitumor mechanism of erythromycin might involve regulating the expression ofc-Myc and p21 (WAF1/Cip1) proteins.