Departement of Developmental Biology and Cancer Research, Institute for Medical Research Israel-Canada, Hadassah Medical School-Hebrew University, Jerusalem, Israel.
PLoS One. 2013;8(1):e54321. doi: 10.1371/journal.pone.0054321. Epub 2013 Jan 14.
Vav1 functions as a signal transducer protein in the hematopoietic system, where it is exclusively expressed. Vav1 was recently implicated in several human cancers, including lung, pancreatic and neuroblasoma. In this study, we analyzed the expression and function of Vav1 in human breast tumors and breast cancer cell lines. Immunohistochemical analysis of primary human breast carcinomas indicated that Vav1 is expressed in 62% of 65 tumors tested and is correlated positively with estrogen receptor expression. Based on published gene profiling of 50 breast cancer cell lines, several Vav1-expressing cell lines were identified. RT-PCR confirmed Vav1 mRNA expression in several of these cell lines, yet no detectable levels of Vav1 protein were observed due to cbl-c proteasomal degradation. We used two of these lines, MCF-7 (Vav1 mRNA negative) and AU565 (Vav1 mRNA positive), to explore the effect of Vav1 expression on breast cell phenotype and function. Vav1 expression had opposite effects on function in these two lines: it reduced proliferation and enhanced cell death in MCF-7 cells but enhanced proliferation in AU565 cells. Consistent with these findings, transcriptome analysis revealed an increase in expression of proliferation-related genes in Vav1-expressing AU565 cells compared to controls, and an increase in apoptosis-related genes in Vav1-expressing MCF-7 cells compared with controls. TUNEL and γ-H2AX foci assays confirmed that expression of Vav1 increased apoptosis in MCF-7 cells but not AU565 cells and shRNA experiments revealed that p53 is required for this pro-apoptotic effect of Vav1 in these cells. These results highlight for the first time the potential role of Vav1 as an oncogenic stress activator in cancer and the p53 dependence of its pro-apoptotic effect in breast cells.
Vav1 在造血系统中作为信号转导蛋白发挥作用,仅在造血系统中表达。Vav1 最近被牵连到几种人类癌症中,包括肺癌、胰腺癌和神经母细胞瘤。在这项研究中,我们分析了 Vav1 在人类乳腺癌肿瘤和乳腺癌细胞系中的表达和功能。对 65 个测试的原发性人类乳腺癌的免疫组织化学分析表明,Vav1 在 62%的肿瘤中表达,并且与雌激素受体表达呈正相关。根据已发表的 50 个乳腺癌细胞系的基因谱分析,确定了几个表达 Vav1 的细胞系。RT-PCR 证实了这些细胞系中的几个细胞系中存在 Vav1 mRNA 表达,但由于 cbl-c 蛋白酶体降解,未观察到可检测水平的 Vav1 蛋白。我们使用这两个细胞系,MCF-7(Vav1 mRNA 阴性)和 AU565(Vav1 mRNA 阳性),探索 Vav1 表达对乳腺细胞表型和功能的影响。Vav1 表达对这两条线的功能有相反的影响:它降低了 MCF-7 细胞的增殖并增强了细胞死亡,但增强了 AU565 细胞的增殖。与这些发现一致,转录组分析显示,与对照相比,表达 Vav1 的 AU565 细胞中增殖相关基因的表达增加,而与对照相比,表达 Vav1 的 MCF-7 细胞中凋亡相关基因的表达增加。TUNEL 和 γ-H2AX 焦点测定法证实,Vav1 的表达增加了 MCF-7 细胞中的细胞凋亡,但不增加 AU565 细胞中的细胞凋亡,并且 shRNA 实验表明,p53 是 Vav1 在这些细胞中促凋亡作用所必需的。这些结果首次强调了 Vav1 作为致癌应激激活物在癌症中的潜在作用,以及其在乳腺细胞中促凋亡作用对 p53 的依赖性。