Department of Bone and Soft Tissue Tumor, National Clinical Research Center for Cancer, Tianjin Medical University Cancer Institute and Hospital, Hexi, Tianjin 300060, P.R. China.
Food and Drug College, Anhui Science and Technology University, Fengyang, Anhui 233100, P.R. China.
Mol Med Rep. 2018 May;17(5):7388-7394. doi: 10.3892/mmr.2018.8746. Epub 2018 Mar 14.
The present study aimed to investigate the role of long non-coding RNA (lncRNA) H19 in the development of osteosarcoma and to determine the underlying mechanism involved. A total of 40 patients with osteosarcoma were selected and the expression level of H19 in tumor tissue and adjacent healthy tissue was detected by reverse transcription‑quantitative polymerase chain reaction. Survival curves were plotted using the Kaplan‑Meier method to investigate the prognostic value of H19 expression level for patients with osteosarcoma. H19 knockdown osteosarcoma cell lines were constructed using small interfering (si)RNA transfection. Cell migration and invasion abilities were measured by Transwell migration and invasion assays, respectively. Western blot analysis was performed to detect the expression levels of phosphatidylinositol 3‑kinase (PI3K), phospho (p)‑PI3K, RAC‑alpha serine/threonine‑protein kinase (AKT), p‑AKT and NF‑κB inhibitor α (IκBα) in osteosarcoma cells transfected with H19 siRNA. Expression level of H19 was significantly elevated in tumor tissue compared with adjacent healthy tissue. Expression level of H19 was positively associated with distant metastasis of osteosarcoma (P<0.01), but not with gender and age. Overall survival of patients with osteosarcoma with high H19 level was significantly shorter compared with patients with low H19 expression (P<0.05). H19 knockdown significantly reduced migration and invasion ability of osteosarcoma cells. Significantly decreased levels of p‑PI3K and p‑AKT, and elevated level of IκBα were observed in H19 knockdown osteosarcoma cells compared with control osteosarcoma cells, while no significant differences in levels of PI3K and AKT were observed. Therefore, the present study demonstrated that knockdown of lncRNA H19 can inhibit migration and invasion of human osteosarcoma cells by inhibiting the nuclear factor-κB pathway.
本研究旨在探讨长链非编码 RNA(lncRNA)H19 在骨肉瘤发展中的作用,并确定涉及的潜在机制。选择 40 例骨肉瘤患者,采用逆转录-定量聚合酶链反应检测肿瘤组织和相邻正常组织中 H19 的表达水平。采用 Kaplan-Meier 法绘制生存曲线,探讨 H19 表达水平对骨肉瘤患者预后的预测价值。采用小干扰(si)RNA 转染构建 H19 敲低骨肉瘤细胞系。通过 Transwell 迁移和侵袭实验分别测量细胞迁移和侵袭能力。采用 Western blot 分析检测转染 H19 siRNA 的骨肉瘤细胞中磷脂酰肌醇 3-激酶(PI3K)、磷酸化(p)-PI3K、RAC-α丝氨酸/苏氨酸-蛋白激酶(AKT)、p-AKT 和核因子-κB 抑制剂α(IκBα)的表达水平。与相邻正常组织相比,肿瘤组织中 H19 的表达水平明显升高。H19 的表达水平与骨肉瘤的远处转移呈正相关(P<0.01),但与性别和年龄无关。H19 高表达的骨肉瘤患者总生存期明显短于 H19 低表达的患者(P<0.05)。H19 敲低显著降低骨肉瘤细胞的迁移和侵袭能力。与对照组骨肉瘤细胞相比,H19 敲低骨肉瘤细胞中 p-PI3K 和 p-AKT 的水平显著降低,IκBα的水平升高,而 PI3K 和 AKT 的水平无显著差异。因此,本研究表明,敲低 lncRNA H19 可通过抑制核因子-κB 通路抑制人骨肉瘤细胞的迁移和侵袭。