Identification of G1-regulated genes in normally cycling human cells.
Identification of G1-regulated genes in normally cycling human cells.
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DOI：
10.1371/journal.pone.0003943
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发表时间：
2008
期刊：
影响因子：
3.7
通讯作者：
Hurt MM
中科院分区：
文献类型：
--
作者：
Beyrouthy MJ;Alexander KE;Baldwin A;Whitfield ML;Bass HW;McGee D;Hurt MM
获得同步细胞群对于细胞周期研究至关重要。诸如撤血清或使用在细胞周期中特定点阻断细胞的药物等方法会改变重新进入细胞周期时的细胞事件。新细胞周期早期 G1 期发生的调控事件可能被忽视。我们使用机器人有丝分裂抖落装置来选择有丝分裂晚期的细胞进行全基因组基因表达研究。进行了两项独立的微阵列实验，其中一项涉及从同步细胞群中每小时分离 RNA，持续几个小时，另一项实验从有丝分裂末期后期到 G1 期每 15 分钟检查一次基因活性。为了验证所研究的细胞群的同步性，我们采用了 BrdU 摄取、FACS 和组蛋白基因活性微阵列分析等方法。我们还检查了应激反应基因的活性。我们的分析能够鉴定出 200 个早期 G1 调控基因，其中许多目前功能未知。我们还通过 qPCR 确认了一组候选基因（fos、atf3 和 tceb）的表达，以进一步验证新鉴定的基因。对自然循环细胞中 G1 期前两个小时的基因组规模表达分析，使得在细胞分裂周期正常进展的细胞中发现了一组独特的 G1 调节基因，其中许多目前具有未知的功能。这组基因可能包含药物开发和人类疾病治疗的未来靶标。
Obtaining synchronous cell populations is essential for cell-cycle studies. Methods such as serum withdrawal or use of drugs which block cells at specific points in the cell cycle alter cellular events upon re-entry into the cell cycle. Regulatory events occurring in early G1 phase of a new cell cycle could have been overlooked. We used a robotic mitotic shake-off apparatus to select cells in late mitosis for genome-wide gene expression studies. Two separate microarray experiments were conducted, one which involved isolation of RNA hourly for several hours from synchronous cell populations, and one experiment which examined gene activity every 15 minutes from late telophase of mitosis into G1 phase. To verify synchrony of the cell populations under study, we utilized methods including BrdU uptake, FACS, and microarray analyses of histone gene activity. We also examined stress response gene activity. Our analysis enabled identification of 200 early G1-regulated genes, many of which currently have unknown functions. We also confirmed the expression of a set of genes candidates (fos, atf3 and tceb) by qPCR to further validate the newly identified genes. Genome-scale expression analyses of the first two hours of G1 in naturally cycling cells enabled the discovery of a unique set of G1-regulated genes, many of which currently have unknown functions, in cells progressing normally through the cell division cycle. This group of genes may contain future targets for drug development and treatment of human disease.
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影响因子：
5.3
作者：
HURT, MM;BOWMAN, TL;MARZLUFF, WF
通讯作者：
MARZLUFF, WF
影响因子：
14.9
作者：
Huang DW;Sherman BT;Tan Q;Kir J;Liu D;Bryant D;Guo Y;Stephens R;Baseler MW;Lane HC;Lempicki RA
通讯作者：
Lempicki RA
DOI：
10.1159/000132871
发表时间：
1989-01-01
期刊：
CYTOGENETICS AND CELL GENETICS
影响因子：
--
作者：
EICHENLAUBRITTER, U;BOLL, I
通讯作者：
BOLL, I
影响因子：
8
作者：
Ji, C;Marnett, LJ;Pietenpol, JA
通讯作者：
Pietenpol, JA
影响因子：
8
作者：
de Belle, I;Huang, RP;Adamson, ED
通讯作者：
Adamson, ED