Targeting programmed cell death ligand 1 by CRISPR/Cas9 in osteosarcoma cells.
Targeting programmed cell death ligand 1 by CRISPR/Cas9 in osteosarcoma cells.
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DOI：
10.18632/oncotarget.16326
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发表时间：
2017-05-02
期刊：
影响因子：
--
通讯作者：
Duan Z
中科院分区：
文献类型：
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作者：
Liao Y;Chen L;Feng Y;Shen J;Gao Y;Cote G;Choy E;Harmon D;Mankin H;Hornicek F;Duan Z
程序性细胞死亡配体1 （PD-L1）是一种在肿瘤细胞上表达的跨膜蛋白，可抑制T细胞介导的免疫反应。靶向PD-L1途径的治疗促进抗肿瘤免疫，并在某些类型的癌症中显示出令人鼓舞的结果。然而，PD-L1在骨肉瘤中的功能和治疗作用在很大程度上仍然未知。在本研究中，我们发现PD-L1蛋白在骨肉瘤细胞系和肿瘤患者的组织微阵列中表达。组织微阵列免疫组化分析显示，PD-L1高表达患者的总生存率和5年生存率明显短于低表达患者。高水平的PD-L1表达也与骨肉瘤患者的转移有关。此外，我们应用聚集规则间隔短回文重复序列(CRISPR)/Cas9系统在骨肉瘤细胞系的DNA水平上靶向PD-L1基因。我们发现，PD-L1的表达可以被CRISPR/Cas9系统有效地破坏，PD-L1的敲低增加了对阿霉素和紫杉醇的药物敏感性。这些结果表明，PD-L1是骨肉瘤的一个独立预后因素，通过CRISPR/Cas9敲除PD-L1可能是治疗骨肉瘤的一种治疗方法。
Programmed cell death ligand 1 (PD-L1) is a transmembrane protein that is expressed on tumor cells that suppresses the T cell-mediated immune response. Therapies targeting the PD-L1 pathway promote anti-tumor immunity and have shown promising results in some types of cancers. However, the functional and therapeutic roles of PD-L1 in osteosarcoma remain largely unknown. In this study, we found that PD-L1 protein was expressed in osteosarcoma cell lines and tissue microarray of patient tumors. Tissue microarray immunohistochemistry analysis showed that the overall and five-year survival rates of patients with high levels of PD-L1 expression were significantly shorter than patients with low levels. High levels of PD-L1 expression were also associated with metastasis in osteosarcoma patients. Furthermore, we applied the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system to target PD-L1 gene at the DNA level in osteosarcoma cell lines. We found that the expression of PD-L1 could be efficiently disrupted by CRISPR/Cas9 system and PD-L1 knockdown increased drug sensitivities for doxorubicin and paclitaxel. These results suggest that PD-L1 is an independent prognostic factor in osteosarcoma and that PD-L1 knockout by CRISPR/Cas9 may be a therapeutic approach for the treatment of osteosarcoma.