Epigenetic silencing of methyl-CpG-binding protein 2 gene affects proliferation, invasion, migration, and apoptosis of human osteosarcoma cells
Epigenetic silencing of methyl-CpG-binding protein 2 gene affects proliferation, invasion, migration, and apoptosis of human osteosarcoma cells
复制标题
甲基CpG结合蛋白2基因的表观遗传沉默影响人骨肉瘤细胞的增殖、侵袭、迁移和凋亡
DOI：
10.1007/s13277-014-2336-8
复制
发表时间：
2014-12-01
期刊：
影响因子：
--
通讯作者：
Guo, Qiao-Nan
中科院分区：
文献类型：
--
作者：
Meng, Gang;Lv, YangFan;Guo, Qiao-Nan
甲基化CpG结合蛋白2（Methyl-CpG-binding protein 2，MeCP 2）是甲基化CpG结合蛋白家族中的一个DNA甲基化相关基因。本研究旨在研究MeCP 2在SaOS 2和U2 OS细胞系中的表观遗传学功能，并探讨MeCP 2基因沉默对骨肉瘤的抗肿瘤作用。本研究采用染色质免疫沉淀法检测MeCP 2与TSSC 3基因的结合活性。RT-PCR和western blot检测LV-MECP 2-RNAi转染后骨肉瘤细胞系中MeCP 2的表达。Transwell侵袭和迁移实验检测细胞的侵袭和迁移能力。流式细胞术检测细胞凋亡情况。还评估了肿瘤大小以确定基因沉默的治疗效果。结果表明，MeCP 2与TSSC 3基因的配合力最高。LV-MECP 2-RNAi能降低肿瘤细胞MeCP 2的表达（P< 0.05）。与对照组相比，LV-MECP 2-RNAi抑制U2 OS和SaOS 2细胞的侵袭和迁移（P< 0.05）。与对照组相比，LV-MECP 2-RNAi可诱导U2 OS和SaOS 2细胞凋亡，并显著抑制细胞增殖（P< 0.05）。与未处理细胞相比，RNAi基因沉默也能显著减小肿瘤体积（P< 0.05）。结论：沉默MeCP 2基因可抑制MeCP 2的表达，抑制肿瘤细胞的迁移、侵袭和增殖，并通过诱导肿瘤细胞凋亡而缩小肿瘤体积。
Methyl-CpG-binding protein 2 (MeCP2) is a DNA methylation-related gene of the methyl-CpG-binding protein family. Here, we investigated the epigenetic function of the MeCP2 in SaOS2 and U2OS cell lines, and explored the antitumor effects of the gene silencing for osteosarcoma. In this study, chromatin immunoprecipitation assay was used to detect MeCP2 binding activity with TSSC3 gene. RT-PCR and western blot assay were used to analyze the MeCP2 expression in osteosarcoma cell lines after transfection with LV-MECP2-RNAi. Transwell invasion and migration assays were used to detect the cell invasion and migration. The cell apoptosis was examined by using the flow cytometry assay. The tumor size was also assessed to determine the therapeutic effects of gene silencing. The results indicated that MeCP2 indicated the highest combining power with TSSC3 gene. LV-MECP2-RNAi could decrease MeCP2 level in tumor cells compared with the untreated cells (P< 0.05). LV-MECP2-RNAi inhibited the U2OS and SaOS2 cells invasion and migration compared with the control cells (P< 0.05). LV-MECP2-RNAi triggered the U2OS and SaOS2 cell apoptosis, and inhibited the cell proliferation significantly compared with the control cells (P< 0.05). The gene silencing of RNAi could also decreased the tumor size significantly compared with untreated cells (P< 0.05). In conclusion, silencing the MeCP2 gene could block the MeCP2 expression and inhibit the tumor cell migration, invasion, and proliferation, and decreases the tumor size by inducing the apoptosis of the tumor cells.