miR-27b-3p通过靶向SSRP1调控骨肉瘤细胞的生长
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常州市第二人民医院骨科,南京医科大学常州医学中心

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常州市科技计划;南京医科大学常州医学中心科研项目


MiR-27b-3p inhibits osteosarcoma cell growth by targeting SSRP1
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The Affiliated Changzhou Second People’s Hospital of Nanjing Medical University

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Changzhou Sci&Tech Program;Subject of Changzhou Medical Center of Nanjing Medical University

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    摘要:

    目的:探究miR-27b-3p对骨肉瘤(osteosarcoma,OS)细胞生长的影响及其潜在的分子机制。方法:①采用实时荧光定量PCR(qRT-PCR)分析miR-27b-3p在骨肉瘤细胞系中的表达。②CCK-8实验、集落形成实验及流式细胞术检测miR-27b-3p以及SSRP1对骨肉瘤细胞生长的影响。③使用在线数据库预测miR-27b-3p的靶基因并进行筛选,使用双荧光素报告酶实验测定miR-27b-3p与结构特异性识别蛋白1(SSRP1)的关系。④干扰效率以及过表达效率使用Western blot实验验证。结果:①miR-27b-3p在骨肉瘤细胞和临床样本中低表达,过表达miR-27b-3p会抑制骨肉瘤细胞的生长。②生物信息学工具和双荧光素报告酶实验证实SSRP1为miR-27b-3p的靶基因,SSRP1表达受miR-27b-3p的直接调控。③干扰SSRP1的表达会抑制骨肉瘤细胞的增殖,促进凋亡。④过表达SSRP1会部分逆转miR-27b-3p对骨肉瘤细胞生长的抑制效应。结论:miR-27b-3p在骨肉瘤细胞中低表达,通过靶向SSRP1从而影响骨肉瘤细胞的生长,证实了miR-27b-3p是骨肉瘤的一个潜在的分子靶点,因此靶向miR-27b-3p/SSRP1轴可能成为骨肉瘤治疗的新策略。

    Abstract:

    Objective: To investigate the effect of miR-27b-3p on osteosarcoma cell growth and its potential molecular mechanism. Methods: ①The expression of miR-27b-3p in osteosarcoma cell lines was analyzed by qRT-PCR. ②CCK-8 assay, colony formation assay and flow cytometry analysis were used to detect the effect of miR-27b-3p and SSRP1 on osteosarcoma cell growth.③The miRNA target prediction database was used to predicted potential target gene of miR-27b-3p, and dual luciferase reporter assays were performed to demonstrate the relationship of miR-27b-3p and its target genes. ④Efficiency of knockdown or overexpression was validated by Western blot analysis. Results: ①miR-27b-3p was lowly expressed in osteosarcoma cell lines and clinical samples, and overexpression of miR-27b-3p led to suppress OS cell growth. ② Bioinformatics analyses and dual-fluorescence reporter assays confirmed that SSRP1 was a target gene of miR-27b-3p, and the expression of SSRP1 can be regulated by miR-27b-3p. ③Silencing of SSRP1 significantly inhibited cell proliferation and increased cell apoptosis in OS. ④Overexpression of SSRP1 partly reversed the inhibitory effect of miR-27b-3p on OS cell growth. Conclusion: miR-27b-3p was downregulated and regulated OS cell growth by targeting SSRP1, this study confirmed that miR-27b-3p was a potential therapeutic target in OS and targeting the miR-27b-3p/SSRP1 axis may become a new therapeutic strategy for the treatment of OS.

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  • 收稿日期:2022-10-08
  • 最后修改日期:2023-02-24
  • 录用日期:2023-04-17
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