Prostate specific membrane antigen knockdown impairs the tumorigenicity of LNCaP prostate cancer cells by inhibiting the phosphatidylinositol 3-kinase/Akt signaling pathway
Prostate specific membrane antigen knockdown impairs the tumorigenicity of LNCaP prostate cancer cells by inhibiting the phosphatidylinositol 3-kinase/Akt signaling pathway
复制标题
前列腺特异性膜抗原敲低通过抑制磷脂酰肌醇 3-激酶/Akt 信号通路损害 LNCaP 前列腺癌细胞的致瘤性
DOI：
10.3760/cma.j.issn.0366-6999.20132707
复制
发表时间：
2014-03-05
影响因子：
6.1
通讯作者：
Huang Hai
中科院分区：
文献类型：
--
作者：
Guo Zhenghui;Lai Yiming;Huang Hai
背景前列腺特异性膜抗原（PSMA）可以促进LNCaP前列腺癌细胞系的生长、迁移和侵袭，但其分子机制尚未明确。本研究采用PSMA基因敲除模型和PI 3 K药理学抑制剂LY 294002在LNCaP前列腺癌细胞中研究PSMA是否作为一种新型的磷脂酰肌醇3-激酶（PI 3 K）/Akt信号通路的调节剂。然后将LNCaP细胞分为干扰组、非干扰组和空白组。我们首先证明了PSMA敲低在我们的LNCaP细胞系中的功效。然后，我们通过Western blotting比较上述三组中有或没有LY 294002处理的PSMA和总/活化Akt的表达。免疫细胞化学染色观察各组细胞内Akt（p-Akt，Ser 473）的变化。结果PSMA基因敲除后，p-Akt（Ser 473）表达水平较未干预组和空白组明显降低，而总Akt（Akt 1/2）表达水平无明显变化。然而，LY 294002给药显著降低了所有三组中p-Akt（Ser 473）的表达。免疫细胞化学结果进一步证实PSMA基因敲除或LY 294002处理与p-Akt（Ser 473）下调有关。结论PI 3 K/Akt信号通路的抑制可能是PSMA基因敲减LNCaP前列腺癌细胞的一种新的分子机制，Akt（Ser 473）可能作为PSMA下游信号靶效应子在该细胞模型中发挥重要作用。
Background Prostate specific membrane antigen (PSMA) can facilitate the growth, migration, and invasion of the LNCaP prostate cancer cell lines, but the underlying molecular mechanisms have not yet been clearly defined. Here, we investigated whether PSMA serves as a novel regulator of the phosphatidylinositol 3-kinase (PI3K)/Akt signaling by employing PSMA knockdown model and PI3K pharmacological inhibitor (LY294002) in LNCaP prostate cancer cells.Methods PSMA knockdown had been stably established by transfecting with lentivirus-mediated siRNA in our previous study. Then, LNCaP cells were divided into interference, non-interference, and blank groups. We first testified the efficacy of PSMA knockdown in our LNCaP cell line. Then, we compared the expression of PSMA and total/activated Akt by Western blotting in the above three groups with or without LY294002 treatment. Furthermore, immunocytochemistry was performed to confirm the changes of activated Akt (p-Akt, Ser473) in groups. Besides, cell proliferation, migration, and cell cycle were measured by CCK-8 assay, Transwell analysis, and Flow cytometry respectively.Results After PSMA knockdown, the level of p-Akt (Ser473) but not of total-Akt (Akt1/2) was significantly decreased when compared with the non-interference and blank groups. However, LY294002 administration significantly reduced the expression of p-Akt (Ser473) in all the three groups. The results of immunocytochemistry further confirmed that PSMA knockdown or LY294002 treatment was associated with p-Akt (Ser473) down-regulation. Decrease of cell proliferation, migration, and survival were also observed upon PSMA knockdown and LY294002 treatment.Conclusions Taken together, our results reveal that PI3K/Akt signaling pathway inhibition may serve as a novel molecular mechanism in LNCaP prostate cancer cells of PSMA knockdown and suggest that Akt (Ser473) may play a critical role as a downstream signaling target effector of PSMA in this cellular model.