长链非编码 RNA MEG3 通过调控 Notch 信号通路抑制骨肉瘤细胞的增殖并促进其凋亡。
LncRNA MEG3 inhibits proliferation and promotes apoptosis of osteosarcoma cells through regulating Notch signaling pathway.
机构信息
Department of Spinal Surgery, The People's Hospital of Rizhao, Rizhao, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Jan;24(2):581-590. doi: 10.26355/eurrev_202001_20034.
OBJECTIVE
To explore the effect of long non-coding ribonucleic acid (lncRNA)-maternally expressed gene 3 (MEG3) on the Notch signaling pathway, and its influences on the proliferation and apoptosis of osteosarcoma MG-63 cells.
MATERIALS AND METHODS
LncRNA MEG3 was overexpressed in osteosarcoma MG-63 cells, and the cells were divided into Blank group, Len-con group, and Len-MEG3 group. The expression level of MEG3 in each group was detected via quantitative Polymerase Chain Reaction (qPCR), the cell proliferation level in each group was detected via Cell Counting Kit-8 (CCK-8) assay, and the apoptosis in each group was detected via Hoechst 33258 staining. Moreover, the content of the inflammatory factors in each group was determined using the Enzyme-Linked Immunosorbent Assay (ELISA), and the expression levels of apoptosis-related proteins and Notch signaling pathway-related proteins were determined through Western blotting.
RESULTS
The expression level of lncRNA MEG3 in Len-MEG3 group was significantly higher than that in the Blank group and Len-con group (p<0.01). The overexpression of lncRNA MEG3 could significantly weaken the proliferation (p<0.01) and enhance the apoptosis of osteosarcoma cells (p<0.01). The overexpression of lncRNA MEG3 could significantly increase the content of the inflammatory factor interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) (p<0.01), and remarkably decrease the content of the anti-inflammatory factor IL-10 in osteosarcoma cells (p<0.01). Besides, the overexpression of lncRNA MEG3 could evidently raise the expression of Caspase3 (p<0.01) and reduce the Bcl-2/Bax expression in osteosarcoma cells (p<0.01). Finally, the overexpression of lncRNA MEG3 could remarkably reduce the protein expressions of Jagged1, Notch1, and NICD1 in osteosarcoma cells (p<0.01).
CONCLUSIONS
The overexpression of lncRNA MEG3 can inhibit the proliferation and promote the apoptosis of osteosarcoma MG-63 cells by suppressing the Notch signaling pathway.
目的
探讨长链非编码 RNA（lncRNA）-母系表达基因 3（MEG3）对 Notch 信号通路的影响及其对骨肉瘤 MG-63 细胞增殖和凋亡的影响。
材料与方法
在骨肉瘤 MG-63 细胞中转染过表达 lncRNA MEG3，将细胞分为空白组、Len-con 组和 Len-MEG3 组。采用实时荧光定量聚合酶链式反应（qPCR）检测各组 MEG3 的表达水平，细胞计数试剂盒（CCK-8）法检测各组细胞的增殖水平，Hoechst 33258 染色法检测各组细胞的凋亡情况。此外，采用酶联免疫吸附试验（ELISA）检测各组细胞中炎症因子的含量，采用 Western blot 检测凋亡相关蛋白和 Notch 信号通路相关蛋白的表达水平。
结果
Len-MEG3 组 lncRNA MEG3 的表达水平明显高于空白组和 Len-con 组（p<0.01）。过表达 lncRNA MEG3 可显著减弱骨肉瘤细胞的增殖（p<0.01），促进细胞凋亡（p<0.01）。过表达 lncRNA MEG3 可显著增加骨肉瘤细胞中炎症因子白细胞介素 6（IL-6）和肿瘤坏死因子-α（TNF-α）的含量（p<0.01），显著降低细胞中抗炎因子白细胞介素 10（IL-10）的含量（p<0.01）。此外，过表达 lncRNA MEG3 可显著上调骨肉瘤细胞中 Caspase3 的表达（p<0.01），下调 Bcl-2/Bax 的表达（p<0.01）。最后，过表达 lncRNA MEG3 可显著降低骨肉瘤细胞中 Jagged1、Notch1 和 NICD1 的蛋白表达（p<0.01）。
结论
过表达 lncRNA MEG3 可通过抑制 Notch 信号通路抑制骨肉瘤 MG-63 细胞的增殖，促进细胞凋亡。
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