中国骨质疏松杂志

LncRNA SNHG16调控miR-214-3p对IL-1β诱导的骨关节炎模型细胞增殖和凋亡的影响

Effect of LncRNA SNHG16 to regulate miR-214-3p on IL-1β-induced cell proliferation and apoptosis of osteoarthritis models

DOI：10.3969/j.issn.1006-7108.2025.12.005

中文关键词: 长链非编码RNA小核仁RNA宿主基因16 微小RNA-214-3p 骨关节炎细胞增殖细胞凋亡

英文关键词:long non coding RNA small nucleolar RNA host gene 16 microRNA-214-3p osteoarthritis cell proliferation cell apoptosis

基金项目:湖北省自然科学基金（2022HBA164）

作者	单位
杨琼1 李健伟2 曾寒1 曹园3*	1.武汉科技大学附属普仁医院骨科,湖北武汉 430000 2.武汉科技大学附属普仁医院手足显微外科,湖北武汉 430000 3.湖北文理学院附属襄阳市中心医院骨科,湖北襄阳 441000

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中文摘要:

目的研究长链非编码RNA小核仁RNA宿主基因16（LncRNA SNHG16）调控微小RNA（miR）-214-3p对白细胞介素-1β（IL-1β）诱导的骨关节炎（OA）模型细胞增殖和凋亡的影响。方法将人软骨细胞CHON-001随机分为NC组、IL-1β组、sh-con组、sh-LncRNA SNHG16组、sh-LncRNA SNHG16+anti-miR-con组、sh-LncRNA SNHG16+anti-miR-214-3p组，每组进行6个重复。除NC组常规培养外，其他组均使用IL-1β诱导OA模型细胞。双荧光素酶报告评估LncRNA SNHG16和miR-214-3p的靶向关系；CCK8、平板克隆、流式细胞仪实验测定各组细胞的存活率、克隆数、凋亡率水平；qRT-PCR实验测定各组细胞中LncRNA SNHG16、miR-214-3p水平；Western blot实验测定各组细胞中PCNA、Bcl-2、Bax、Cleaved Caspase-3蛋白水平。结果双荧光素酶报告发现，WT-LncRNA SNHG16+miR-214-3p mimic组比WT-LncRNA SNHG16+mimic-con组细胞的荧光素酶活性低（P＜0.05）。与NC组相比，IL-1β组细胞的存活率、克隆数、miR-214-3p、PCNA、Bcl-2表达减少，凋亡率、LncRNA SNHG16、Bax、Cleaved Caspase-3表达增多（P＜0.05）；与IL-1β组、sh-con组相比，sh-LncRNA SNHG16组细胞的存活率、克隆数、miR-214-3p、PCNA、Bcl-2表达增多，凋亡率、LncRNA SNHG16、Bax、Cleaved Caspase-3表达减少（P＜0.05）；与sh-LncRNA SNHG16组、sh-LncRNA SNHG16+anti-miR-con组相比，sh-LncRNA SNHG16+anti-miR-214-3p组细胞的存活率、克隆数、miR-214-3p、PCNA、Bcl-2表达减少，凋亡率、Bax、Cleaved Caspase-3表达增多（P＜0.05）。结论下调LncRNA SNHG16可能通过靶向上调miR-214-3p的表达来抑制IL-1β诱导的OA模型细胞的凋亡，促进其增殖。

英文摘要:

Objective To investigate the effect of long non-coding RNA small nucleolar RNA host gene 16 (LncRNA SNHG16) on the proliferation and apoptosis of osteoarthritis (OA) chondrocytes induced by interleukin-1β (IL-1β) through regulation of microRNA (miR-214-3p). Methods Human chondrocyte CHON-001 was randomly divided into NC group, IL-1β group, sh-con group, sh-LncRNA SNHG16 group, sh-LncRNA SNHG16+anti-miR-con group, sh-LncRNA SNHG16+anti- miR-214-3p group, each group was subjected to 6 replicates. Except for routine culture in the NC group, all the other groups used IL-1β to induce OA model cells. Dual luciferase assay was used to evaluate the targeting relationship between LncRNA SNHG16 and miR-214-3p; CCK8, plate cloning and flow cytometry experiments determine the survival rate, cloning number and apoptosis rate levels of each group of cells; qRT-PCR experiments were used to determine the levels of LncRNA SNHG16 and miR-214-3p in each group of cells; Western blot experiments were used to determine the levels of PCNA, Bcl-2, Bax, and Cleaved Caspase-3 proteins in each group of cells. Results The dual luciferase report found that the cells in the WT-LncRNA SNHG16+miR-214-3p mimic group had lower luciferase activity than the WT-LncRNA SNHG16+mimic-con group (P<0.05). Compared with the NC group, the survival rate, clone number, miR-214-3p, PCNA, and Bcl-2 expression of cells in the IL-1β group decreased, and the apoptosis rate, LncRNA SNHG16, Bax, and Cleaved Caspase-3 expression increased (P<0.05). Compared with the IL-1β group and sh-con group, the survival rate, clone number, miR-214-3p, PCNA, and Bcl-2 expression increased, apoptosis rate, LncRNA SNHG16 group, Bax, Cleaved Caspase-3 expression was reduced (P<0.05). Compared with the sh-LncRNA SNHG16 group and sh-LncRNA SNHG16+anti-miR-con group, the survival rate, clone number, miR-214-3p, PCNA of the sh-LncRNA SNHG16+anti-miR-214-3p group, Bcl-2 expression decreased, and apoptosis rate, Bax, and Cleaved Caspase-3 expression increased (P<0.05). Conclusion Downregulation of LncRNA SNHG16 may inhibit IL-1β-induced OA model cells apoptosis and promote proliferation by targeting upregulation of miR-214-3p.

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