长非编码 RNA H19 通过 Wnt/β-catenin 信号通路调控胶质瘤的发生发展。
Long non-coding RNA H19 regulates the development of gliomas through the Wnt/β-catenin signaling pathway.
机构信息
Neurosurgery, First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China.
出版信息
Eur Rev Med Pharmacol Sci. 2019 May;23(10):4243-4253. doi: 10.26355/eurrev_201905_17929.
OBJECTIVE
We investigated the effects of long non-coding RNA (lncRNA) H19 on glioma cell proliferation, invasion, migration, and apoptosis and the underlying mechanisms.
PATIENTS AND METHODS
H19 expression in glioma tissues, para-carcinoma tissues, and glioma cell lines was analyzed by Real-time polymerase chain reaction (RT-PCR). After transfecting U251 and U87MG cells with siRNA-H19, cell proliferation was detected by the cell counting kit-8 (CCK8) assay. Invasion and migration were detected by a transwell assay; cell cycle distribution and apoptosis were measured by flow cytometry analysis; Dvl2, GSK-3β, cyclin D1, and β-catenin expressions were detected by RT-PCR and Western blotting.
RESULTS
H19 expression in glioma tissues was higher than that in para-carcinoma tissues and associated with poor prognosis in glioma patients. Cell proliferation, invasion, and migration significantly decreased, the percentage of glioma cells in G0/G1 significantly increased, the percentage of glioma cells in the S phase significantly decreased, and apoptosis significantly increased in U251 and U87MG cells transfected with siRNA-H19 compared to those in the siRNA-NC group. Downregulation of H19 decreased DVL2, cyclin D1, and β-catenin expression and increased GSK-3β expression. The inhibitory effects of downregulation of H19 on glioma cell proliferation, invasion, and migration were reversed by SKL2001 via the activation of the Wnt/β-catenin signal pathway, which was further enhanced by inhibition of the Wnt/β-catenin signal pathway by XAV939.
CONCLUSIONS
H19 was overexpressed in glioma tissues and glioma cell lines. Downregulation of H19 inhibited cell proliferation, invasion, and migration, arrested cell cycle progression in the G0/G1 phase, and induced cell apoptosis by restraining activation of the Wnt/β-catenin signaling pathway in glioma cells. Therefore, H19 is a potential therapeutic target for glioma therapy.
目的
研究长链非编码 RNA（lncRNA）H19 对神经胶质瘤细胞增殖、侵袭、迁移和凋亡的影响及其作用机制。
方法
采用实时聚合酶链反应（RT-PCR）检测神经胶质瘤组织、癌旁组织和神经胶质瘤细胞系中 H19 的表达。用 siRNA-H19 转染 U251 和 U87MG 细胞后，用细胞计数试剂盒-8（CCK8）检测细胞增殖。通过 Transwell 检测侵袭和迁移；通过流式细胞术分析细胞周期分布和凋亡；通过 RT-PCR 和 Western blot 检测 Dvl2、GSK-3β、cyclin D1 和β-catenin 的表达。
结果
神经胶质瘤组织中 H19 的表达高于癌旁组织，与神经胶质瘤患者的不良预后相关。与 siRNA-NC 组相比，转染 siRNA-H19 的 U251 和 U87MG 细胞的细胞增殖、侵袭和迁移明显减少，G0/G1 期神经胶质瘤细胞比例明显增加，S 期神经胶质瘤细胞比例明显减少，细胞凋亡明显增加。下调 H19 降低了 DVL2、cyclin D1 和β-catenin 的表达，增加了 GSK-3β 的表达。通过激活 Wnt/β-catenin 信号通路，SKL2001 逆转了下调 H19 对神经胶质瘤细胞增殖、侵袭和迁移的抑制作用，通过 XAV939 抑制 Wnt/β-catenin 信号通路进一步增强了这种抑制作用。
结论
H19 在神经胶质瘤组织和神经胶质瘤细胞系中过度表达。下调 H19 抑制神经胶质瘤细胞增殖、侵袭和迁移，通过抑制 Wnt/β-catenin 信号通路使细胞周期停滞在 G0/G1 期，并诱导细胞凋亡。因此，H19 是神经胶质瘤治疗的潜在治疗靶点。
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