Dual blockade of PKA and NF-κB inhibits H2 relaxin-mediated castrate-resistant growth of prostate cancer sublines and induces apoptosis.
Dual blockade of PKA and NF-κB inhibits H2 relaxin-mediated castrate-resistant growth of prostate cancer sublines and induces apoptosis.
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DOI：
10.1007/s12672-011-0076-4
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发表时间：
2011-08
影响因子：
3
通讯作者：
de Vere White, Ralph W
中科院分区：
文献类型：
--
作者：
Vinall, Ruth L;Mahaffey, Christopher M;Davis, Ryan R;Luo, Zunping;Gandour-Edwards, Regina;Ghosh, Paramita M;Tepper, Clifford G;de Vere White, Ralph W
我们先前证明了H2松弛素（RLN 2）通过PI 3 K/Akt/β-连环蛋白介导的雄激素受体（AR）通路的激活促进前列腺癌（CaP）细胞的去势抵抗（CR）生长。由于抑制该途径仅导致CR生长减少约50%，因此本研究的目标是确定有助于CR生长的其他RLN 2激活途径。基于下一代测序（NGS）的转录组和基因本体（GO）分析比较了用RLN 2稳定转染的LNCaP（LNCaP-RLN 2）与LNCaP-载体，确定了与细胞增殖相关的基因的差异表达（12.7%的差异表达基因），包括与cAMP/PKA和NFκB途径相关的基因。随后的分子分析证实cAMP/PKA和NFκB通路在促进H2松弛素介导的CaP细胞CR生长中起作用。PKA的抑制减弱了RLN 2介导的AR活性，抑制了增殖，并引起了小但显著的凋亡增加。通过抑制PKA和Akt联合抑制PKA和NFκB信号传导通路可诱导显著的细胞凋亡，并显著降低克隆形成潜力，优于多西他赛（CR CaP的标准治疗）。组织微阵列（TMA）的免疫组织化学（IHC）分析结合多光谱定量成像，比较了BPH、PIN和CaP患者中的RLN 2水平，确定了CaP与BPH相比，RLN 2显著上调（p=0.002）。综合数据表明，RLN 2过表达在CaP患者中很常见，并为CaP细胞提供了生长优势。RLN 2诱导的CR生长的几乎完全抑制可以通过同时阻断两种途径来实现。
We previously demonstrated H2 relaxin (RLN2) facilitates castrate resistant (CR) growth of prostate cancer (CaP) cells through PI3K/Akt/β-catenin-mediated activation of the androgen receptor (AR) pathway. As inhibition of this pathway caused only ~50% reduction in CR growth, the goal of the current study was to identify additional RLN2-activated pathways that contribute to CR growth. Next-generation sequencing (NGS)-based transcriptome and gene ontology (GO) analyses comparing LNCaP stably transfected with RLN2 (LNCaP-RLN2) versus LNCaP-vector identified differential expression of genes associated with cell proliferation (12.7% of differentially expressed genes), including genes associated with the cAMP/PKA and NFκB pathways. Subsequent molecular analyses confirmed that the cAMP/PKA and NFκB pathways play a role in facilitating H2 relaxin-mediated CR growth of CaP cells. Inhibition of PKA attenuated RLN2-mediated AR activity, inhibited proliferation and caused a small but significant increase in apoptosis. Combined inhibition of the PKA and NFκB signaling pathways via inhibition of PKA and Akt induced significant apoptosis and dramatically reduced clonogenic potential, outperforming docetaxel, the standard of care treatment for CR CaP. Immunohistochemical (IHC) analysis of tissue microarrays (TMA) in combination with multispectral quantitative imaging comparing RLN2 levels in patients with BPH, PIN and CaP determined that RLN2 is significantly upregulated in CaP vs BPH (p=0.002). The combined data indicate RLN2 overexpression is frequent in CaP patients and provides a growth advantage to CaP cells. A near complete inhibition of RLN2-induced CR growth can be achieved by simultaneous blockade of both pathways.