cDNA microarray analysis of invasive and tumorigenic phenotypes in a breast cancer model
cDNA microarray analysis of invasive and tumorigenic phenotypes in a breast cancer model
复制标题
DOI：
10.1038/labinvest.3700044
复制
发表时间：
2004-03-01
影响因子：
5
通讯作者：
Sapi, E
中科院分区：
文献类型：
--
作者：
Kluger, HM;Kluger, Y;Sapi, E
FMS癌基因编码巨噬细胞集落刺激因子受体(CSF1R)，这是一种跨膜酪氨酸激酶受体，在乳腺癌中异常表达。将野生型CSF1R导入HC11乳腺上皮细胞(HC11-CSF1R)，使其具有体外局部侵袭和体内成瘤能力。用突变的CSF1R在Tyr-721自动磷酸化位点(HC11-CSF1R-721)表达Phe，可产生一种缺乏转移能力但保持局部侵袭性的表型。相反，在Tyr-807点突变的CSF1R基因突变的HC11细胞(HC11-CSF1R-807)保持其转移能力，但不具有局部侵袭性。我们的目的是利用4.6K的基因芯片，确定野生型CSF1R转染HC11后哪些基因有差异表达，以及自磷酸化位点突变对基因表达的影响。将来自HC11、HC11-CSF1R-721和HC11-CSF1R-807的互补DNA分别与HC11-CSF1R杂交。样品聚类采用主成分光谱分析方法，并结合前向正交化过程。经方差分析鉴定差异表达基因。Northern blotting证实MAP激酶磷酸酶-1、WDNM1(细胞外蛋白水解酶抑制物)、trp-2(肿瘤相关钙信号转导蛋白-2)、IV型前胶原、分泌型白蛋白抑制物、早烯基化SNARE蛋白Ykt6、铜蓝蛋白和伴侣蛋白10等基因与肿瘤的侵袭和转移无关。我们已经成功地确定了与侵袭性表型或肿瘤发生有关的基因。这些基因为进一步研究转移进展和局部侵袭性提供了基础，可以作为治疗靶点进行评估。
The fms oncogene encodes the macrophage colony-stimulating factor receptor (CSF1R), a transmembrane tyrosine kinase receptor, which is abnormally expressed in breast cancer. Transfection of wild-type CSF1R into HC11 mammary epithelial cells (HC11-CSF1R) renders the transfectants capable of in vitro local invasion and in vivo tumorigenesis. Transfection with CSF1R mutated to express phe at the tyr-721 autophosphorylation site (HC11-CSF1R-721) creates a phenotype that lacks metastastic competence but maintains local invasiveness. Conversely, HC11 cells transfected with CSF1R mutated at tyr-807 (HC11-CSF1R-807) retain their metastatic competence, but are not locally invasive. Our aims were to determine which genes were differentially expressed with transfection of HC11 with wild-type CSF1R, and to determine the effect of mutation at the autophosphorylation sites on gene expression, using 4.6 K cDNA microarrays. Complementary DNA from HC11, HC11-CSF1R-721 and HC11-CSF1R-807 were each hybridized together with HC11-CSF1R on individual arrays. A principal component spectral method combined with prenormallization procedures was used for sample clustering. Differentially expressed genes were identified by the analysis of variance. Confirmation by Northern blotting was performed for MAP kinase phosphatase-1, WDNM1 (extracellular proteinase inhibitor), Trop 2 (tumor-associated calcium signal transducer-2), procollagen type IV alpha, secretory leukoprotease inhibitor, prenylated snare protein Ykt6, ceruloplasmin and chaperonin 10. Many of these genes have not previously been associated with tumor invasion and metastasis. We have successfully identified genes that can be linked to the invasive phenotypes or to tumorigenesis. These genes provide a basis for further studies of metastatic progression and local invasiveness, and can be evaluated as therapeutic targets.