Cryptochrome 2 (CRY2) Suppresses Proliferation and Migration and Regulates Clock Gene Network in Osteosarcoma Cells.
Cryptochrome 2 (CRY2) Suppresses Proliferation and Migration and Regulates Clock Gene Network in Osteosarcoma Cells.
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DOI：
10.12659/msm.908596
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发表时间：
2018-06-07
期刊：
影响因子：
--
通讯作者：
Hong Y
中科院分区：
文献类型：
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作者：
Yu Y;Li Y;Zhou L;Yang G;Wang M;Hong Y
昼夜节律紊乱是人类潜在的癌症风险因素。然而，时钟基因隐花色素2（Cryptochrome 2，CRP 2）在骨肉瘤（osteosarcoma，OS）中的作用尚不清楚。为了评估HOS骨肉瘤细胞中HOS 2的潜在作用，建立了HOS 2沉默细胞系。此外，我们研究了CCK-8，集落形成，迁移实验，流式细胞术，在体外HOS细胞的影响，敲低p53 2。HOS-2基因敲低可促进HOS-OS细胞增殖和迁移。我们使用细胞周期测定来显示，G12敲低增加了S期细胞群体，减少了G1期细胞群体。Western blot分析显示，P53基因敲低降低了P53的表达，增加了c-myc和cyclin D1的表达。同时，敲低JNK 2增加了细胞外信号调节激酶（ERK）1/2的磷酸化，但不改变c-Jun N末端激酶（JNK）和P38的磷酸化。RP 12基因敲低还可通过诱导细胞周期进程、促进丝裂原活化蛋白激酶（MAPK）和Wnt/β-catenin信号通路，增加基质金属蛋白酶（MMP）-2和β-catenin的表达，促进OS细胞增殖和迁移。虽然之前还不清楚BMAL 2的表达是否会影响时钟基因网络中其他时钟基因的表达，但我们的研究结果表明，BMAL 2的敲低显著增加了BMAL 1，Period（PER）1，PER 2，BMAL 1和CLOCK的mRNA表达。我们的研究结果表明，在OS，其功能可能涉及下游基因和其他昼夜节律基因的抑癌基因。
Circadian disruption is a potential cancer risk factor in humans. However, the role of the clock gene, cryptochrome 2 (CRY2), in osteosarcoma (OS) is still not clear. To evaluate the potential role of CRY2 in HOS osteosarcoma cells, CRY2-silenced cell lines were established. Furthermore, we investigated the effect of CRY2 knockdown on HOS cells by CCK-8, colony formation, migration assay, and flow cytometry, in vitro. CRY2 knockdown promoted HOS OS cell proliferation and migration. We used a cell cycle assay to show that CRY2 knockdown increased the S phase cell population and reduced the G1 phase cell population. Western blot analyses showed that CRY2 knockdown decreased P53 expression and increased expression of c-myc and cyclin D1. Simultaneously, CRY2 knockdown increased the phosphorylation of extracellular signal-regulated kinase (ERK) 1/2, but did not change the phosphorylation of c-Jun N terminal kinase (JNK) and P38. CRY2 knockdown also increased the expression of matrix metalloproteinase (MMP)-2 and β-catenin, and increased OS cell proliferation and migration by inducing cell cycle progression and promoting mitogen-activated protein kinase (MAPK) and Wnt/β-catenin signaling pathways. Although it has previously been unclear whether the expression of CRY2 affects the expression of other clock genes in the clock gene network, our results show that knockdown of CRY2 significantly increased the mRNA expression of CRY1, Period (PER) 1, PER2, BMAL1, and CLOCK. Our results suggest that CRY2 may be an anti-oncogene in OS, whose functions involve both downstream genes and other circadian genes.