Phenotypic effects of the circadian gene Cryptochrome 2 on cancer-related pathways.
Phenotypic effects of the circadian gene Cryptochrome 2 on cancer-related pathways.
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DOI：
10.1186/1471-2407-10-110
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发表时间：
2010-03-24
期刊：
影响因子：
3.8
通讯作者：
Zhu Y
中科院分区：
文献类型：
--
作者：
Hoffman AE;Zheng T;Ba Y;Stevens RG;Yi CH;Leaderer D;Zhu Y
昼夜节律基因作为重要的转录调节因子继续受到关注，有可能影响多种生物途径，包括许多与癌症相关的过程。核心昼夜节律基因隐花色素 2 (CRY2) 对于正确的昼夜节律计时至关重要，并且是昼夜节律反馈环路负臂的关键组成部分。因此，CRY2 的异常表达可能会影响致癌过程，从而影响癌症易感性。我们使用小干扰寡核苷酸 (siRNA) 沉默乳腺癌细胞系 (MCF-7) 中的 CRY2，并测量 CRY2 敲低对许多癌症相关参数的影响。使用流式细胞术在暴露和未暴露于诱变剂攻击的细胞中测量 CRY2 敲低 (CRY2-) 和正常 (CRY2+) 细胞群的细胞周期分布、细胞活力和凋亡反应。使用单细胞凝胶电泳（彗星）测定法测量 DNA 损伤积累，并使用 Olive 尾矩对损伤进行量化，该方法考虑了 DNA 远离细胞核迁移的数量和距离，表明 DNA 链断裂。通过全基因组微阵列测量癌症相关转录本的表达变化。使用学生 t 检验进行统计比较，并使用错误发现率校正对从微阵列获得的 P 值进行调整以进行多重比较，以获得每次观察的调整后的 Q 值。彗星试验结果表明，暴露于相同剂量的化学诱变剂后，CRY2-细胞比CRY2+细胞积累了更多未修复的DNA损伤（P = 0.040），这表明CRY2可能对DNA修复很重要。此外，许多与 DNA 损伤修复相关的转录本在 CRY2 沉默后表现出表达改变。其中包括 BCCIP (Q = 0.002)、BCL2 (Q = 0.049)、CCND1 (Q = 0.009)、CDKN1A (Q < 0.001)、GADD45A (Q = 0.002)、HERC5 (Q < 0.001)、MCM5 (Q = 0.042)、PPP1R15A (Q < 0.001)、SUMO1 (Q < 0.001) 和 UBA1 (Q = 0.023)。然而，没有检测到 CRY2 敲低对细胞周期分布、响应诱变剂攻击的细胞周期检查点或细胞凋亡反应的显着影响。总的来说，这些数据表明 CRY2 在调节 DNA 损伤修复和维持基因组稳定性方面发挥有限但潜在的重要作用。未来的研究可能集中于确定 CRY2 调节已知与致癌相关的转录物表达的机制。
Circadian genes continue to gain attention as important transcriptional regulators with the potential to influence a variety of biological pathways, including many cancer-related processes. The core circadian gene cryptochrome 2 (CRY2) is essential for proper circadian timing, and is a key component of the negative arm of the circadian feedback loop. As such, aberrant expression of CRY2 may influence carcinogenic processes and thereby impact cancer susceptibility. We silenced CRY2 in breast cancer cell lines (MCF-7) using small-interfering oligos (siRNA) and measured the impact of CRY2 knockdown on a number of cancer-relevant parameters. Cell cycle distribution, cell viability, and apoptotic response were measured in CRY2 knockdown (CRY2-) and normal (CRY2+) cell populations using flow cytometry in cells with and without exposure to a mutagen challenge. DNA damage accumulation was measured using the single cell gel electrophoresis (comet) assay, and damage was quantified using the Olive tail moment, which considers the amount and distance of DNA migration away from the nucleus, indicative of DNA strand breaks. Expression changes in cancer-relevant transcripts were measured by whole genome microarray. The Student's t-test was used for statistical comparisons, and P-values obtained from the microarray were adjusted for multiple comparisons using the false discovery rate correction, in order to obtain an adjusted Q-value for each observation. The comet assay results indicated that upon exposure to the same dose of chemical mutagen, CRY2- cells accumulate significantly more unrepaired DNA damage than CRY2+ cells (P = 0.040), suggesting that CRY2 may be important for DNA repair. In addition, a number of transcripts with relevance for DNA damage repair displayed altered expression following CRY2 silencing. These included BCCIP (Q = 0.002), BCL2 (Q = 0.049), CCND1 (Q = 0.009), CDKN1A (Q < 0.001), GADD45A (Q = 0.002), HERC5 (Q < 0.001), MCM5 (Q = 0.042), PPP1R15A (Q < 0.001), SUMO1 (Q < 0.001), and UBA1 (Q = 0.023). However, no significant influence of CRY2 knockdown on cell cycle distributions, cell cycle checkpoints in response to mutagen challenge, or apoptotic response was detected. In total, these data suggest a limited, but potentially important role for CRY2 in the regulation of DNA damage repair and the maintenance of genomic stability. Future investigations may focus on identifying the mechanisms by which CRY2 may regulate the expression of transcripts with known relevance for carcinogenesis.
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影响因子：
64.8
作者：
Bartkova, J;Horejsi, Z;Bartek, J
通讯作者：
Bartek, J
影响因子：
--
作者：
Xiang, Shulin;Coffelt, Seth B;Mao, Lulu;Yuan, Lin;Cheng, Qi;Hill, Steven M
通讯作者：
Hill, Steven M
DOI：
10.1073/pnas.0409897102
发表时间：
2005-03-01
影响因子：
11.1
作者：
Gorbacheva, VY;Kondratov, RV;Antoch, MP
通讯作者：
Antoch, MP
影响因子：
11.2
作者：
Gauger, MA;Sancar, A
通讯作者：
Sancar, A
影响因子：
56.9
作者：
Matsuo, T;Yamaguchi, S;Okamura, H
通讯作者：
Okamura, H