Centre de recherche du Centre Hospitalier de l'Université de Montréal (CR/CHUM)/Institut du cancer de Montréal, Montréal, Canada.
J Ovarian Res. 2009 Apr 14;2:4. doi: 10.1186/1757-2215-2-4.
We previously observed the over-expression of BMP-2 in primary cultures of epithelial ovarian cancer (EOC) cells as compared to normal epithelial cells based on Affymetrix microarray profiling 1. Here we investigate the effect of BMP-2 on several parameters of ovarian cancer tumorigenesis using the TOV-2223, TOV-1946 and TOV-112D EOC cell lines.
We treated each EOC cell line with recombinant BMP-2 and assayed various parameters associated with tumorigenesis. More specifically, cell signaling events induced by BMP-2 treatment were investigated by western-blot using anti-phosphospecific antibodies. Induction of Id1, Snail and Smad6 mRNA expression was investigated by real time RT-PCR. The ability of cells to migrate was tested using the scratch assay. Cell-cell adhesion was analyzed by the ability of cells to form spheroids. We also investigated BMP-2 expression in tissue samples from a series of EOC patients.
Treatment of these cell lines with recombinant BMP-2 induced a rapid phosphorylation of Smad1/5/8 and Erk MAPKs. Increased expression of Id1, Smad6 and Snail mRNAs was also observed. Only in the TOV-2223 cell line were these signaling events accompanied by an alteration in cell proliferation. We also observed that BMP-2 efficiently increased the motility of all three cell lines. In contrast, BMP-2 treatment decreased the ability of TOV-1946 and TOV-112D cell lines to form spheroids indicating an inhibition of cell-cell adhesion. The expression of BMP-2 in tumor tissues from patients was inversely correlated with survival.
These results suggest that EOC cell secretion of BMP-2 in the tumor environment contributes to a modification of tumor cell behavior through a change in motility and adherence. We also show that BMP-2 expression in tumor tissues is associated with a poorer prognosis for ovarian cancer patients.
我们之前通过 Affymetrix 微阵列分析发现，与正常上皮细胞相比，上皮性卵巢癌（EOC）细胞的原代培养中 BMP-2 过度表达。在这里，我们使用 TOV-2223、TOV-1946 和 TOV-112D EOC 细胞系研究了 BMP-2 对卵巢癌发生的几个参数的影响。
我们用重组 BMP-2 处理每种 EOC 细胞系，并检测与肿瘤发生相关的各种参数。更具体地说，使用抗磷酸化特异性抗体通过 Western blot 研究 BMP-2 处理诱导的细胞信号转导事件。通过实时 RT-PCR 研究 Id1、Snail 和 Smad6 mRNA 表达的诱导。使用划痕实验测试细胞迁移能力。通过细胞形成球体的能力分析细胞-细胞黏附。我们还研究了一系列 EOC 患者组织样本中的 BMP-2 表达。
用重组 BMP-2 处理这些细胞系会迅速诱导 Smad1/5/8 和 Erk MAPKs 的磷酸化。还观察到 Id1、Smad6 和 Snail mRNA 的表达增加。只有在 TOV-2223 细胞系中，这些信号事件伴随着细胞增殖的改变。我们还观察到 BMP-2 有效地增加了所有三种细胞系的运动能力。相比之下，BMP-2 处理降低了 TOV-1946 和 TOV-112D 细胞系形成球体的能力，表明细胞-细胞黏附的抑制。患者肿瘤组织中 BMP-2 的表达与生存呈负相关。
这些结果表明，EOC 细胞在肿瘤微环境中分泌的 BMP-2 通过改变运动和粘附来改变肿瘤细胞的行为。我们还表明，肿瘤组织中 BMP-2 的表达与卵巢癌患者的预后较差相关。
