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慢病毒介导的 shRNA 敲低人骨肉瘤细胞中的 MED19 通过诱导 G0/G1 期细胞周期阻滞抑制细胞增殖。

Knockdown of MED19 by lentivirus-mediated shRNA in human osteosarcoma cells inhibits cell proliferation by inducing cell cycle arrest in the G0/G1 phase.

机构信息

Department of Orthopaedics, The First Hospital of Jilin University, Changchun, China.

出版信息

Oncol Res. 2011;19(5):193-201. doi: 10.3727/096504011x12970940207760.

Abstract

MED 19 is a subunit of the mediator complex, which is a coactivator of RNA polymerase II and also interacts with the downstream coding region of many genes. However, the role of MED19 in osteosarcoma is unknown. In the present study, we applied lentivirus-mediated short hairpin RNA (shRNA)-triggered RNA interference to downregulate MED19 expression in human osteosarcoma SaOS-2 and U2OS cells. Knockdown of MED19 expression was confirmed by real-time PCR and Western blot. It was found that silencing of MED19 resulted in decreased cell viability, colony formation capacity, and DNA synthesis ability in both cells, as well as the G0/G1 phase cell cycle arrest. These results implied that MED19 played an important role in cell growth and cell cycle progression of human osteosarcoma cells. MED19 may be an attractive candidate for the therapeutic target in osteosarcoma.

摘要

MED19 是中介体复合物的一个亚基,它是 RNA 聚合酶 II 的共激活因子,也与许多基因的下游编码区相互作用。然而,MED19 在骨肉瘤中的作用尚不清楚。在本研究中,我们应用慢病毒介导的短发夹 RNA(shRNA)触发 RNA 干扰来下调人骨肉瘤 SaOS-2 和 U2OS 细胞中的 MED19 表达。通过实时 PCR 和 Western blot 验证了 MED19 表达的下调。结果发现,沉默 MED19 表达导致两种细胞的细胞活力、集落形成能力和 DNA 合成能力下降,以及 G0/G1 期细胞周期停滞。这些结果表明,MED19 在人骨肉瘤细胞的生长和细胞周期进程中发挥着重要作用。MED19 可能是骨肉瘤治疗靶点的一个有吸引力的候选物。

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