SNAI1 is critical for the aggressiveness of prostate cancer cells with low E-cadherin.
SNAI1 is critical for the aggressiveness of prostate cancer cells with low E-cadherin.
复制标题
DOI：
10.1186/1476-4598-13-37
复制
发表时间：
2014-02-24
期刊：
影响因子：
37.3
通讯作者：
Agarwal R
中科院分区：
文献类型：
--
作者：
Deep G;Jain AK;Ramteke A;Ting H;Vijendra KC;Gangar SC;Agarwal C;Agarwal R
更好地了解前列腺癌发生的分子机制，以制定新的针对前列腺癌（PCA）的预防和治疗策略是必要的，前列腺癌是男性死亡的主要原因。在这里，我们研究了两个上皮-间质转化（EMT）调节剂，粘附连接蛋白E-钙粘蛋白及其转录抑制因子SNAI 1，在调节PCA细胞的侵略性的作用。在细胞培养和裸鼠异种移植模型中，在MTT和克隆形成测定中比较具有稳定敲低的E-钙粘蛋白的人前列腺癌PC 3细胞（ShEC-PC 3）和相应的对照细胞（Sh-PC 3）的生长速率。前列腺素测定法分析ShEC-PC 3和Sh-PC 3细胞的干细胞。Western blotting和免疫组织化学（IHC）用于研究E-cadherin和SNAI 1敲低后蛋白表达的变化。采用小干扰RNA（siRNA）技术敲低ShEC-PC 3细胞中SNAI 1蛋白的表达。ShEC-PC 3细胞在细胞培养和裸鼠体内的增殖率均高于Sh-PC 3细胞。ShEC-PC 3细胞还形成了更大且数量显着增加的前列腺球，这表明随着E-钙粘蛋白敲低，干细胞样群体增加。此外，与Sh-PC 3前列腺球相比，在血清和附着存在下，ShEC-PC 3前列腺球崩解产生更大质量的增殖细胞。免疫印迹/IHC分析显示，E-钙粘蛋白敲低增加了细胞培养物和异种移植组织中干性（CD 44、切割的Notch 1和Egr-1）和EMT（波形蛋白、pSrc-tyr 416、整合素β3、β-连环蛋白和NF-κB）的调节因子/生物标志物的表达。几种骨转移相关分子，即CXCR 4、uPA、RANKL和RunX 2在ShEC-PC 3细胞中的表达也增加。重要的是，我们观察到SNAI 1在ShEC-PC 3细胞的细胞质和细胞核组分、前列腺球和异种移植组织中的表达显著增加。SNAI 1基因的特异性siRNA敲除可显著抑制前列腺球的形成、克隆形成和侵袭能力，并降低PSrc-tyr 416、总Src和CD 44的表达水平。RWPE-1、WPE 1-NA 22、WPE 1-NB 14和DU-145细胞的表征进一步证实了低E-钙粘蛋白与较高的SNAI 1表达和前列腺球形成相关。总之，这些结果表明，E-钙粘蛋白损失促进SNAI 1表达，控制PCA细胞的侵略性。
A better molecular understanding of prostate carcinogenesis is warranted to devise novel targeted preventive and therapeutic strategies against prostate cancer (PCA), a major cause of mortality among men. Here, we examined the role of two epithelial-to-mesenchymal transition (EMT) regulators, the adherens junction protein E-cadherin and its transcriptional repressor SNAI1, in regulating the aggressiveness of PCA cells. The growth rate of human prostate carcinoma PC3 cells with stable knock-down of E-cadherin (ShEC-PC3) and respective control cells (Sh-PC3) was compared in MTT and clonogenic assays in cell culture and in nude mouse xenograft model in vivo. Stemness of ShEC-PC3 and Sh-PC3 cells was analyzed in prostasphere assay. Western blotting and immunohistochemistry (IHC) were used to study protein expression changes following E-cadherin and SNAI1 knock-down. Small interfering RNA (siRNA) technique was employed to knock- down SNAI1 protein expression in ShEC-PC3 cells. ShEC-PC3 cells exerted higher proliferation rate both in cell culture and in athymic nude mice compared to Sh-PC3 cells. ShEC-PC3 cells also formed larger and a significantly higher number of prostaspheres suggesting an increase in the stem cell-like population with E-cadherin knock-down. Also, ShEC-PC3 prostaspheres disintegration, in the presence of serum and attachment, generated a bigger mass of proliferating cells as compared to Sh-PC3 prostaspheres. Immunoblotting/IHC analyses showed that E-cadherin knock-down increases the expression of regulators/biomarkers for stemness (CD44, cleaved Notch1 and Egr-1) and EMT (Vimentin, pSrc-tyr416, Integrin β3, β-catenin, and NF-κB) in cell culture and xenograft tissues. The expression of several bone metastasis related molecules namely CXCR4, uPA, RANKL and RunX2 was also increased in ShEC-PC3 cells. Importantly, we observed a remarkable increase in SNAI1 expression in cytoplasmic and nuclear fractions, prostaspheres and xenograft tissues of ShEC-PC3 cells. Furthermore, SNAI1 knock-down by specific siRNA strongly inhibited the prostasphere formation, clonogenicity and invasiveness, and decreased the level of pSrc-tyr416, total Src and CD44 in ShEC-PC3 cells. Characterization of RWPE-1, WPE1-NA22, WPE1-NB14 and DU-145 cells further confirmed that low E-cadherin is associated with higher SNAI1 expression and prostasphere formation. Together, these results suggest that E-cadherin loss promotes SNAI1 expression that controls the aggressiveness of PCA cells.
登录
查看更多内容
DOI：
10.1016/j.urolonc.2007.05.010
发表时间：
2007-09-01
影响因子：
2.7
作者：
Hadaschik, Boris A.;Gleave, Martin E.
通讯作者：
Gleave, Martin E.
影响因子：
6.1
作者：
Baygi, Modjtaba Emadi;Soheili, Zahra Soheila;Schulz, Wolfgang A.
通讯作者：
Schulz, Wolfgang A.
影响因子：
64.5
作者：
Gupta PB;Onder TT;Jiang G;Tao K;Kuperwasser C;Weinberg RA;Lander ES
通讯作者：
Lander ES
影响因子：
8
作者：
Baritaki, S.;Chapman, A.;Bonavida, B.
通讯作者：
Bonavida, B.
影响因子：
11.5
作者：
Balic, Marija;Lin, Henry;Cote, Richard J.
通讯作者：
Cote, Richard J.