PRL-3 suppresses c-Fos and integrin α2 expression in ovarian cancer cells.
PRL-3 suppresses c-Fos and integrin α2 expression in ovarian cancer cells.
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DOI：
10.1186/1471-2407-13-80
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发表时间：
2013-02-18
期刊：
影响因子：
3.8
通讯作者：
Zeng Q
中科院分区：
文献类型：
--
作者：
Liu H;Al-aidaroos AQ;Wang H;Guo K;Li J;Zhang HF;Zeng Q
再生肝磷酸酶-3(PRL-3)是一种蛋白酪氨酸磷酸酶，在多种人类肿瘤中高表达，与肿瘤进展和肿瘤转移密切相关。然而，PRL-3促进癌细胞迁移、侵袭和转移的机制还不是很清楚。在本研究中，我们探讨了PRL-3在卵巢癌进展中的作用及其分子机制。免疫组织化学方法检测卵巢癌组织芯片上PRL-3蛋白的表达。利用短发夹状RNA(ShRNA)构建稳定的PRL-3缺失细胞系。用Transwell法和Matrigel法分析这些细胞的迁移和侵袭能力。免疫印迹和免疫荧光检测PRL-3消融细胞和对照细胞的蛋白水平和分布。苏木精-伊红染色和透射电子显微镜观察细胞形态。最后，将PRL-3消融的细胞和对照细胞注射到裸鼠体内，进行移植瘤的致瘤性分析。在135例卵巢癌患者中，有26例(19%)PRL-3表达升高，而在正常卵巢组织中未见表达(0/14)。与对照亲本A2780细胞相比，稳定缺失PRL-3导致A2780卵巢癌细胞的迁移能力和侵袭活性降低。此外，经PRL-3处理的细胞还表现出扁平的形态和延长的片状脂膜。为了解决与PRL-3下调相关的表型改变的可能的分子基础，我们评估了参与细胞-基质黏附的各种蛋白质的表达谱。去除Prl-3可显著提高细胞表面受体整合素α2的核糖核酸和蛋白质水平，但不能显著提高其异源结合伙伴整合素β1的表达水平。抑制PrL-3还与上调巴西林的表达和磷酸化有关。在这些α-3基因敲除的细胞中，整合素PRL2的转录激活剂c-fos的表达和激活明显增加。此外，强制表达eGFP-PRL-3可抑制A2780细胞整合素α-2和c-fos的表达。值得注意的是，使用异种移植瘤模型，我们在体内观察到A2780 PRL-3基因敲除细胞的致瘤性大大降低。这些结果表明，PRL-3在卵巢癌的发生和维持恶性表型中起重要作用。Prl-3可抑制c-fos对整合素α-2信号的转录调控。我们的结果有力地支持了PRL-3在卵巢癌进展中作为一个有前景的治疗靶点和潜在的早期生物标记物的作用。
Phosphatase of regenerating liver-3 (PRL-3), a protein tyrosine phosphatase, is highly expressed in multiple human cancers and strongly implicated in tumor progression and cancer metastasis. However, the mechanisms by which PRL-3 promotes cancer cell migration, invasion, and metastasis are not very well understood. In this study, we investigated the contribution and molecular mechanisms of PRL-3 in ovarian cancer progression. PRL-3 protein expression was detected on ovarian cancer tissue microarrays using immunohistochemistry. Stable PRL-3 depleted cell lines were generated using short hairpin RNA (shRNA) constructs. The migration and invasion potential of these cells were analyzed using Transwell and Matrigel assays, respectively. Immunoblotting and immunofluorescence were used to detect protein levels and distribution in PRL-3-ablated cells and the control cells. Cell morphology was observed with hematoxylin-eosin staining and transmission electron microscopy. Finally, PRL-3-ablated and control cells were injected into nude mice for xenograft tumorigenicity assays. Elevated PRL-3 expression was detected in 19% (26 out of 135) of human ovarian cancer patient samples, but not in normal ovary tissues (0 out of 14). Stable depletion of PRL-3 in A2780 ovarian cancer cells resulted in decreased migration ability and invasion activity compared with control parental A2780 cells. In addition, PRL-3-ablated cells also exhibited flattened morphology and extended lamellipodia. To address the possible molecular basis for the altered phenotypes associated with PRL-3 down-regulation, we assessed the expression profiles of various proteins involved in cell-matrix adhesion. Depletion of PRL-3 dramatically enhanced both RNA and protein levels of the cell surface receptor integrin α2, but not its heterologous binding partner integrin β1. Inhibition of PRL-3 also correlated with elevated expression and phosphorylation of paxillin. A pronounced increase in the expression and activation of c-fos, a transcriptional activator of integrin α2, was observed in these PRL-3 knock-down cells. Moreover, forced expression of EGFP-PRL-3 resulted in the suppression of both integrin α2 and c-fos expression in A2780 cells. Significantly, using a xenograft tumor model, we observed a greatly reduced tumorigenicity of A2780 PRL-3 knock-down cells in vivo. These results suggest that PRL-3 plays a critical role in ovarian cancer tumorigenicity and maintaining the malignant phenotype. PRL-3 may inhibit c-fos transcriptional regulation of integrin α2 signaling. Our results strongly support a role for PRL-3 as a promising therapeutic target and potential early biomarker in ovarian cancer progression.
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