| 赵轶男,袁 志,毕 龙,郝 赋,牛志霞,孙 强.LncRNA MEG3通过抑制Wnt/β-catenin信号通路对骨关节炎软骨细胞增殖和凋亡的影响[J].,2019,19(24):4617-4623 |
| LncRNA MEG3通过抑制Wnt/β-catenin信号通路对骨关节炎软骨细胞增殖和凋亡的影响 |
| LncRNA MEG3 Regulates the Osteoarthritis Chondrocyte Proliferation and Apoptosis through Inhibiting Wnt/β-catenin Signaling Pathway |
| 投稿时间：2019-06-05 修订日期：2019-06-29 |
| DOI：10.13241/j.cnki.pmb.2019.24.004 |
| 中文关键词: lncRNA MEG3 骨关节炎 Wnt/β-catenin 增殖 凋亡 |
| 英文关键词: LncRNA MEG3 Osteoarthritis Wnt/β-catenin Proliferation Apoptosis |
| 基金项目:国家自然科学基金项目(2016YFC1100304) |
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| 中文摘要: |
| 摘要 目的：探讨长链非编码RNA(Long non-coding RNA，LncRNA)人母系表达基因(Maternally expressed gene 3，MEG3)对骨关节炎(osteoarthritis，OA)软骨细胞增殖和凋亡的影响及作用机制。方法：选取我院住院的OA患者和半月板损伤患者各40例，采用RT-PCR检测两组患者软骨组织和细胞中MEG3的表达。在OA软骨细胞中，转染siRNA- MEG3(si- MEG3)或过表达MEG3的慢病毒载体(LV- MEG3)，采用CCK8法检测细胞增殖情况，流式细胞仪检测细胞周期和凋亡情况，RT-PCR和western blot检测PCNA、Dvl2、GSK-3β、cyclinD1和β-catenin mRNA和蛋白表达。结果：OA患者膝关节软骨组织中MEG3的表达水平明显低于半月板损伤患者软骨组织(P<0.05)，同时OA软骨细胞中MEG3的表达水平明显低于正常软骨细胞(P<0.05)。OA软骨细胞转染si- MEG3后，细胞增殖能力和PCNA表达明显下降(P均<0.05)，G0/G1期细胞比例明显升高(P<0.05)，S期细胞比例明显下降(P<0.05)，细胞凋亡率明显增加(P均<0.05)。低表达MEG3能够显著降低Dvl2、GSK-3β、cyclinD1和β-catenin mRNA和蛋白表达水平(P均<0.05)，增加GSK-3β mRNA和蛋白表达水平(P均<0.05)。在OA软骨细胞转染LV- MEG3后，细胞增殖能力和PCNA表达明显升高(P均<0.05)，G0/G1期细胞比例明显下降(P<0.05)，S期细胞比例明显增加(P<0.05)，细胞凋亡率明显减少(P均<0.05)。高表达MEG3能够显著增加OA软骨细胞中Dvl2、GSK-3β、cyclinD1和β-catenin mRNA和蛋白表达水平(P均<0.05)，降低GSK-3β mRNA和蛋白表达(P均<0.05)。同时，采用XAV939阻滞Wnt/β-catenin信号通路能够显著逆转过表达MEG3对OA软骨细胞增殖和凋亡的影响。结论：MEG3在OA患者和软骨细胞中均显著低表达，并能够通过阻滞Wnt/β-catenin信号通路激活影响OA软骨细胞增殖和凋亡。MEG3可能成为OA治疗的重要靶分子。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effects of Long non-coding RNA (lncRNA) Maternally expressed gene 3 (MEG3) on osteoarthritis (OA) chondrocyte proliferation and apoptosis, and the underlying mechanisms. Methods: 40 patients with OA and 40 patients with meniscus injury in our hospital were selected. MEG3 expression in OA cartilage tissues and cells were analyzed by real-time polymerase chain reaction(RT-PCR). After transfecting OA chondrocytes with si- MEG3 or LV- MEG3, cell proliferation was detected by the cell counting kit-8(CCK8) assay, cell cycle distribution and apoptosis were measured by flow cytometry analysis, and PCNA, Dvl2, GSK-3β, cyclinD1 and β-catenin were detected by RT-PCR and western blotting. Results: The expression level of MEG3 in OA cartilage tissues was lower than that in patients with meniscus injury(P<0.05) and the expression level MEG3 in OA chondrocytes was lower than that in normal chondrocytes(P<0.05). Cell proliferation, and PCNA expression decreased significantly (P<0.05), the percentage of OA chondrocytes in G0/G1 significantly increased(P<0.05), the percentage of OA chondrocytes in the S phase significantly decreased(P<0.05), and apoptosis significantly increased(P<0.05) in OA chondrocytes transfected with si- MEG3 compared to those in the si-NC group. Down regulation of MEG3 decreased DVL2, cyclin D1, and β-catenin expression(P<0.05) and increased GSK-3β expression(P<0.05). Cell proliferation, and PCNA expression increased significantly(P<0.05), the percentage of OA chondrocytes in G0/G1 significantly decreased(P<0.05), the percentage of OA chondrocytes in the S phase significantly increased(P<0.05), and apoptosis significantly decreased(P<0.05) in OA chondrocytes transfected with LV- MEG3 compared to those in the LV-NC group. Up-regulation of MEG3 increased DVL2, cyclin D1, and β-catenin expression(P<0.05) and decreased GSK-3β expression(P<0.05). Meanwhile, the effects of overexpression MEG3 on OA chondrocyte proliferation and apoptosis were reversed by inhibition of the Wnt/β-catenin signal pathway by XAV939. Conclusion: MEG3 expression was down-regulated in both OA patients and chondrocytes, which could affect the proliferation and apoptosis in OA chondrocytes by blocking the activation of Wnt/β-catenin signaling pathway. MEG3 may become an important target molecule for OA treatment. |
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