Effect of CXCR4 on Apoptosis in Osteosarcoma Cells via the PI3K/Akt/NF-κβ Signaling Pathway
Effect of CXCR4 on Apoptosis in Osteosarcoma Cells via the PI3K/Akt/NF-κβ Signaling Pathway
复制标题
CXCR4 通过 PI3K/Akt/NF-β 信号通路对骨肉瘤细胞凋亡的影响
DOI：
10.1159/000489593
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Bian, Zhenyu
中科院分区：
文献类型：
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作者：
Jiang, Chunming;Ma, Shenglin;Bian, Zhenyu
背景/目标：骨肉瘤是最常见的原发性骨恶性肿瘤，起源于间充质来源的原始转化细胞，其发病率和死亡率在世界范围内呈上升趋势。以往的研究发现骨肉瘤细胞的凋亡是促进骨肉瘤进展的有效途径，而CXCR 4已被证实与多种肿瘤的进展和转移有关。方法：BrdU法检测转染CXCR 4 shRNA和对照组细胞的增殖情况。流式细胞术检测细胞凋亡。Western blot检测凋亡蛋白表达水平。采用比色法检测Caspase活性。构建NF-κβ启动子荧光素酶报告质粒，检测骨肉瘤细胞中CXCR 4下调后NF-κβ信号的激活。通过检测骨肉瘤细胞中NF-κβ信号相关蛋白的表达和活化情况，探讨CXCR 4下调后Akt与NF-κβ信号的关系。结果如下：CXCR 4的下调可显著降低骨肉瘤细胞的增殖，同时显著增加细胞凋亡和凋亡蛋白的表达水平。此外，下调CXCR 4诱导的细胞凋亡在骨肉瘤细胞中依赖于caspase。CXCR 4下调可通过抑制PI 3 K/Akt/NF-κβ信号通路诱导细胞凋亡。此外，内质网应激（ERS）激活参与细胞凋亡诱导的CXCR 4下调。部分ERS相关蛋白的敲除和CXCR 4的下调可显著抑制细胞凋亡和凋亡蛋白的表达水平。结论：因此，下调CXCR 4表达可通过抑制PI 3 K/Akt/NF-κβ信号通路诱导人骨肉瘤细胞凋亡，提示CXCR 4对骨肉瘤的临床治疗具有重要意义。
Background/Aims: Osteosarcoma, the most common primary bone malignancy, arises from primitive transformed cells of mesenchymal origin with the worldwide increasing morbidity and mortality. Previous studies found apoptosis of osteosarcoma cells was essential for an effective manner to improve the progress of osteosarcoma, and CXCR4 has been demonstrated to be relevant with various tumor progress and metastasis. Methods: The proliferation of cells transfected with CXCR4 shRNA and control shRNA were measured by BrdU assay. Apoptosis was detected by flow cytometry. Apoptotic protein expression levels were detected by Western blot. Caspase activity was detected by Colorimetric Assay Kits using microplate reader. Activation of NF-κβ signaling after CXCR4 down-regulation in osteosarcoma cells was examined by constructing NF-κβ promoter luciferase reporter plasmid. The expression and activation of NF-κβ Signaling relevant protein were analyzed to investigate the relationship between Akt and NF-κβ signaling after the down-regulation of CXCR4 in osteosarcoma cells. Results: Down-regulation of CXCR4 significantly reduced the cell proliferation, while remarkably increased the cell apoptosis and apoptotic protein expression levels in osteosarcoma cells. Furthermore, down-regulation of CXCR4 induced cell apoptosis was caspase dependent in osteosarcoma cells. This study also showed CXCR4 down-regulation induced apoptosis through inhibiting PI3K/Akt/NF-κβ signaling pathway. In addition, endoplasmic reticulum stress (ERS) activation was involved in cell apoptosis induced down-regulation of CXCR4. Knockdown of partial ERS relevant proteins followed down-regulation of CXCR4 significantly inhibited cell apoptosis and the apoptotic protein expression levels. Conclusions: Taken together, the results demonstrated that down-regulation of CXCR4 could induce apoptosis of human osteosarcoma cells through inhibiting PI3K/Akt/NF-κβ signaling pathway, indicating that CXCR4 could be vital for the clinical therapy of osteosarcoma.