Inhibition of phosphodiesterase-4 (PDE4) activity triggers luminal apoptosis and AKT dephosphorylation in a 3-D colonic-crypt model.
Inhibition of phosphodiesterase-4 (PDE4) activity triggers luminal apoptosis and AKT dephosphorylation in a 3-D colonic-crypt model.
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DOI：
10.1186/1476-4598-11-46
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发表时间：
2012-07-25
期刊：
影响因子：
37.3
通讯作者：
Shirasawa S
中科院分区：
文献类型：
--
作者：
Tsunoda T;Ota T;Fujimoto T;Doi K;Tanaka Y;Yoshida Y;Ogawa M;Matsuzaki H;Hamabashiri M;Tyson DR;Kuroki M;Miyamoto S;Shirasawa S
我们之前建立了一个三维（3-D）结肠隐窝模型使用HKe3细胞，这是人类结直肠癌（CRC）HCT 116细胞与致癌KRAS的破坏，并揭示了致癌KRAS在抑制细胞凋亡和破坏细胞极性的关键作用;然而，KRAS诱导这些3-D特异性生物学变化的分子机制仍有待阐明。在该模型中被致癌KRAS上调的基因中，我们专注于磷酸二酯酶4B（PDE4B），在基因表达分析的公共数据集中，发现其表达水平在来自CRC患者的临床肿瘤样品中高于来自健康对照的那些。PDE4B2在其他PDE4亚型中特异性过表达，HKe3细胞中致癌KRAS的再表达导致PDE4B过表达。此外，使用咯利普兰抑制PDE 4催化活性通过诱导ZO-1（紧密连接标记物）和E-钙粘蛋白（粘附连接标记物）的顶端组装以及通过增加管腔中caspase-3（凋亡标记物）的活性，将HCT 116细胞的解体恢复为上皮细胞极性的正常生理状态。值得注意的是，咯利普兰减少了AKT磷酸化，已知AKT磷酸化与管腔形成和CRC发展的破坏有关。使用PDE4B2-shRNA也获得了类似的结果。此外，在基因表达分析的公开数据集中，发现PDE4B mRNA表达增加与复发性CRC相关。这些结果共同表明，PDE4 B被致癌KRAS上调，并且PDE4催化活性的抑制可以诱导CRC中的上皮细胞极性和腔细胞凋亡，从而突出了我们的3-D培养（3DC）模型用于在3-D微环境中KRAS诱导的CRC发展的效用。事实上，使用这个模型，我们发现PDE4B是一个有前途的候选人的治疗目标，以及预后的分子标志物在CRC。进一步阐明PDE4B2在3DC中的信号传导网络将提供对体内CRC的更好理解。
We previously established a three-dimensional (3-D) colonic crypt model using HKe3 cells which are human colorectal cancer (CRC) HCT116 cells with a disruption in oncogenic KRAS, and revealed the crucial roles of oncogenic KRAS both in inhibition of apoptosis and in disruption of cell polarity; however, the molecular mechanism of KRAS-induced these 3-D specific biological changes remains to be elucidated. Among the genes that were upregulated by oncogenic KRAS in this model, we focused on the phosphodiesterase 4B (PDE4B) of which expression levels were found to be higher in clinical tumor samples from CRC patients in comparison to those from healthy control in the public datasets of gene expression analysis. PDE4B2 was specifically overexpressed among other PDE4 isoforms, and re-expression of oncogenic KRAS in HKe3 cells resulted in PDE4B overexpression. Furthermore, the inhibition of PDE4 catalytic activity using rolipram reverted the disorganization of HCT116 cells into the normal physiologic state of the epithelial cell polarity by inducing the apical assembly of ZO-1 (a tight junction marker) and E-cadherin (an adherens junction marker) and by increasing the activity of caspase-3 (an apoptosis marker) in luminal cavities. Notably, rolipram reduced the AKT phosphorylation, which is known to be associated with the disruption of luminal cavity formation and CRC development. Similar results were also obtained using PDE4B2-shRNAs. In addition, increased expression of PDE4B mRNA was found to be correlated with relapsed CRC in a public datasets of gene expression analysis. These results collectively suggested that PDE4B is upregulated by oncogenic KRAS, and also that the inhibition of PDE4 catalytic activity can induce both epithelial cell polarity and luminal apoptosis in CRC, thus highlighting the utility of our 3-D culture (3 DC) model for the KRAS-induced development of CRC in 3-D microenvironment. Indeed, using this model, we found that PDE4B is a promising candidate for a therapeutic target as well as prognostic molecular marker in CRC. Further elucidation of the signaling network of PDE4B2 in 3 DC would provide a better understanding of CRC in vivo.
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影响因子：
56.9
作者：
Golub, TR;Slonim, DK;Lander, ES
通讯作者：
Lander, ES
影响因子：
4.8
作者：
Debnath, J;Muthuswamy, SK;Brugge, JS
通讯作者：
Brugge, JS
影响因子：
--
作者：
Kenny, Hilary A.;Dogan, Songuel;Zillhardt, Marion;Mitra, Anirban K.;Yamada, S. Diane;Krausz, Thomas;Lengyel, Ernst
通讯作者：
Lengyel, Ernst
影响因子：
11.5
作者：
Hong, Yi;Ho, Kok Sun;Cheah, Peh Yean
通讯作者：
Cheah, Peh Yean
DOI：
10.1073/pnas.86.15.6007
发表时间：
1989-08-01
影响因子：
11.1
作者：
MANGOOKARIM, R;UCHIC, M;GRANTHAM, JJ
通讯作者：
GRANTHAM, JJ