ms-275对骨肉瘤细胞reck基因表达的影响研究word论文.docxVIP

MS-275 对骨肉瘤细胞 RECK 基因表达的影响研究中 文 摘 要目的观察组蛋白去乙酰化酶抑制剂恩替司他（Entinostat，MS-275）对骨肉瘤细 胞 RECK 基因表达的影响。方法体外培养骨肉瘤细胞系 MG-63，并根据不同的实验目的分为空白对照组和 不同浓度 MS-275 处理组。其中对照组仅给予 0.1% DMSO 处理；MS-275 干预组 采用 0.1，1.0，3.0μmol/L MS-275 作用 MG63 细胞 24～72h。采用 WST-1 法检测 MS-275 处理后 MG-63 细胞的增殖情况。提取细胞总 RNA 和蛋白，RT-PCR 检 测 RECK mRNA 和蛋白的表达情况。同时，采用荧光共振能量转移法检测基质 金属蛋白酶（MMP）-9 的酶活性。结果1.WST-1 结果显示，在 0.1～3μmol/L 浓度范围内，MS-275 可显著抑制 MG-63细胞增殖，其中 3μmol/L MS-275 对 MG-63 细胞生长抑制率达 46%。2.RT-PCR 结果显示：RECK 基因在 MG-63 细胞中表达较低。经 0.1～3μmol/L MS-275 处理后，RECK mRNA 水平随着 MS-275 浓度的增高而递增。Western blot 结果也显示，不同浓度 MS-275 也能提高 MG-63 细胞中 RECK 蛋白的表 达水平。3. 酶活性结果显示，MG-63 细胞上清液中 MMP-9 酶活性水平较高，经 0.1～3μmol/L MS-275 处理后，其酶活性随着浓度的增高而降低。其中 3μmol/L2MS-275 可将其酶活性降低至 43%。结论MS-275 能上调 RECK mRNA 和蛋白表达水平，最终影响 MMP-9 的酶活性 而发挥抗肿瘤作用。关键词恩替司他；RECK；骨肉瘤；基质金属蛋白酶 9；3Effect of MS-275 on RECK gene expression in osteosarcoma cellsAbstractObjectiveTo observe the effect of histone deacetylase inhibitors Entinostat (MS-275) on RECK gene expression in osteosarcoma cells.MethodsOsteosarcoma cell line MG-63 was cultured in vitro and divided into control group and different concentrations of MS-275 groups. For the control group, cells weretreated by 0.1% DMSO；For the MS-275 groups, cells were treated by 0.1, 1.0, 3.0μmol/L MS-275 for 24~72h.The multiplication of MG-63 cells were detected by WST-1 assay. Total cellular RNA and protein was extracted, and expression of RECK mRNA and protein were detected by RT-PCR. Meanwhile, enzyme activity of Matrix metalloproteinase-9 (MMP-9) was detected by foerster resonance energy transfer assay.ResultsWST-1 results showed that 0.1~3umol/L MS-275 significantly inhibited the proliferation of MG-63 cells, and the inhibiting rate reached to 46% when 3μmol/L MS-275 was administrated.RT-PCR results showed, RECK gene was lowly expressed in MG-63 cells. After treatment of 0.1~3μmol/L MS-275, the RECK mRNA increased. Western blot results also showed, different concentration MS-275 could increase the expression of RECK protein in MG-