Silencing of protein kinase D2 induces glioma cell senescence via p53-dependent and -independent pathways.
Silencing of protein kinase D2 induces glioma cell senescence via p53-dependent and -independent pathways.
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DOI：
10.1093/neuonc/not303
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发表时间：
2014-07
期刊：
影响因子：
15.9
通讯作者：
Sattler W
中科院分区：
文献类型：
--
作者：
Bernhart E;Damm S;Heffeter P;Wintersperger A;Asslaber M;Frank S;Hammer A;Strohmaier H;DeVaney T;Mrfka M;Eder H;Windpassinger C;Ireson CR;Mischel PS;Berger W;Sattler W
多形性胶质母细胞瘤（GBM）是一种高度侵袭性的中枢神经系统肿瘤，患者预后不良。异常的蛋白激酶C（PKC）信号转导与胶质瘤的发生有关，PKC激活的蛋白激酶D（PRKD）家族的成员PRKD 2被鉴定为体外和体内GBM生长的介质。用不同的胶质瘤细胞系建立PRKD 2沉默和对胶质瘤细胞增殖的药理学抑制的结果。利用Western印迹、衰老测定、免疫共沉淀、荧光激活细胞分选、定量PCR和免疫荧光显微术来分析下游信号传导。PRKD 2的RNA干扰（21-mer siRNA）和药理学抑制（CRT 0066101）可显著抑制p53 wt（U87 MG、A172和原代GBM 2）和p53 mut（GM 133、T98 G、U251和原代Gli 25）神经胶质瘤细胞的增殖。在异种移植实验中，PRKD 2沉默显著延迟了U87 MG细胞的肿瘤生长。p53 wt和p53 mut细胞中的PRKD 2沉默与G1中衰老和细胞周期停滞的典型标志相关。响应于PRKD 2沉默的减弱的AKT/PKB磷酸化是在p53 wt和p53 mut GBM细胞中进行的常见观察。p53 wt细胞中PRKD 2敲低诱导p53、p21和p27表达上调，CDK 2和/或CDK 4磷酸化降低，视网膜母细胞瘤蛋白（pRb）磷酸化不足，以及E2 F1转录降低。在p53 mut GM 133和原代Gli 25细胞中，PRKD 2沉默增加了p27和p15，减少了E2 F1转录，但不影响pRb磷酸化。PRKD 2沉默通过p53依赖性和非依赖性途径诱导胶质瘤细胞衰老。
Glioblastoma multiforme (GBM) is a highly aggressive tumor of the central nervous system with a dismal prognosis for affected patients. Aberrant protein kinase C (PKC) signaling has been implicated in gliomagenesis, and a member of the PKC-activated protein kinase D (PRKD) family, PRKD2, was identified as mediator of GBM growth in vitro and in vivo. The outcome of PRKD2 silencing and pharmacological inhibition on glioma cell proliferation was established with different glioma cell lines. Western blotting, senescence assays, co-immunoprecipitation, fluorescence activated cell sorting, quantitative PCR, and immunofluorescence microscopy were utilized to analyze downstream signaling. RNA-interference (21-mer siRNA) and pharmacological inhibition (CRT0066101) of PRKD2 profoundly inhibited proliferation of p53wt (U87MG, A172, and primary GBM2), and p53mut (GM133, T98G, U251, and primary Gli25) glioma cells. In a xenograft experiment, PRKD2 silencing significantly delayed tumor growth of U87MG cells. PRKD2 silencing in p53wt and p53mut cells was associated with typical hallmarks of senescence and cell cycle arrest in G1. Attenuated AKT/PKB phosphorylation in response to PRKD2 silencing was a common observation made in p53wt and p53mut GBM cells. PRKD2 knockdown in p53wt cells induced upregulation of p53, p21, and p27 expression, decreased phosphorylation of CDK2 and/or CDK4, hypophosphorylation of retinoblastoma protein (pRb), and reduced transcription of E2F1. In p53mut GM133 and primary Gli25 cells, PRKD2 silencing increased p27 and p15 and reduced E2F1 transcription but did not affect pRb phosphorylation. PRKD2 silencing induces glioma cell senescence via p53-dependent and -independent pathways.
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影响因子：
15.9
作者：
Azoitei, Ninel;Kleger, Alexander;Seufferlein, Thomas
通讯作者：
Seufferlein, Thomas
DOI：
10.1073/pnas.92.20.9363
发表时间：
1995-09-26
影响因子：
11.1
作者：
DIMRI, GP;LEE, XH;CAMPISI, J
通讯作者：
CAMPISI, J
影响因子：
10.5
作者：
Kim, Youngmi;Kim, Eunhee;Rich, Jeremy N.
通讯作者：
Rich, Jeremy N.
影响因子：
37.3
作者：
Krakstad C;Chekenya M
通讯作者：
Chekenya M
影响因子：
15.9
作者：
Guersel, Demirkan B.;Connell-Albert, Yvette S.;Reilly, Karlyne M.
通讯作者：
Reilly, Karlyne M.