ST8SIA6-AS1对骨肉瘤细胞增殖和转移的影响及其分子机制

Influence of ST8SIA6-AS1 on proliferation and metastasis of osteosarcoma cells and its molecular mechanism

  • 摘要:
    目的 探讨ST8SIA6-AS1对骨肉瘤细胞增殖和转移的影响及其分子机制。
    方法 选取37例骨肉瘤患者的瘤组织及瘤旁组织,采用实时荧光定量聚合酶链反应(qRT-PCR)检测ST8SIA6-AS1、miR-223-3p的表达水平。将骨肉瘤细胞U2OS分为si-ST8SIA6-AS1组、si-NC组、miR-223-3p mimic组、miR-NC组、si-ST8SIA6-AS1+miR-223-3p inhibitor组、si-ST8SIA6-AS1+anti-miR-NC组。采用四甲基偶氮唑盐比色法(MTT)检测细胞增殖抑制率; 采用克隆形成实验检测细胞克隆形成数; 采用划痕实验检测细胞划痕愈合率; 采用Transwell检测细胞迁移数; 采用双荧光素酶报告实验检测ST8SIA6-AS1与miR-223-3p的靶向关系。
    结果 与瘤旁组织相比,骨肉瘤组织中ST8SIA6-AS1表达水平升高, miR-223-3p表达水平降低,差异有统计学意义(P < 0.05)。沉默ST8SIA6-AS1或过表达miR-223-3p后, U2OS细胞增殖抑制率升高, U2OS细胞克隆形成数和迁移数减少, U2OS细胞划痕愈合率降低,差异均有统计学意义(P < 0.05)。ST8SIA6-AS1可靶向调控miR-223-3p; 抑制miR-223-3p可逆转沉默ST8SIA6-AS1对U2OS的抑制作用。
    结论 ST8SIA6-AS1在骨肉瘤中高表达,沉默ST8SIA6-AS1可通过调控miR-223-3p抑制骨肉瘤U2OS细胞增殖和迁移。

     

    Abstract:
    Objective To explore the influence of ST8SIA6-AS1 on proliferation and metastasis of osteosarcoma cells and its molecular mechanism.
    Methods The tumor tissues and adjacent tissues of 37 patients with osteosarcoma were selected, and the expression levels of ST8SIA6-AS1 and miR-223-3p were detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). The osteosarcoma cells U2OS were divided into si-ST8SIA6-AS1 group, si-NC group, miR-223-3p mimic group, miR-NC group, si-ST8SIA6-AS1+miR-223-3p inhibitor group, and si-ST8SIA6-AS1+anti-miR-NC group. Methyl thiazolyl tetrazolium (MTT) assay was used to detect in hibitory rate of cell proliferation; clone formation test was used to detect formation number of cell clone; scratch test was used to detect healing rate of cell scratch; the transwell test was used to detect cell migration number; dual luciferase report test was used to detect the targeting relationship between ST8SIA6-AS1 and miR-223-3p.
    Results Compared with the adjacent tissues, the expression level of ST8SIA6-AS1 in osteosarcoma tissue was significantly increased, and the expression level of miR-223-3p was significantly decreased (P < 0.05). After silencing ST8SIA6-AS1 or overexpression of miR-223-3p, the inhibitory rate ofU2OS cell proliferation was significantly increased, formation number of U2OS cell clone and migration number were significantly reduced, and healing rate of U2OS cell scratch was significantly reduced (P < 0.05). ST8SIA6-AS1 was able to target and regulate miR-223-3p; inhibiting miR-223-3p was able to reverse the inhibitory effect of silencing ST8SIA6-AS1 on U2OS.
    Conclusion ST8SIA6-AS1 is highly expressed in osteosarcoma, and silencing ST8SIA6-AS1 can inhibit the proliferation and migration of osteosarcoma U2OS cells by regulating miR-223-3p.

     

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