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c-Ha-ras癌基因转化的人乳腺上皮细胞的癌蛋白表达及形态学表型

Oncoprotein expression and morphological phenotypes of human breast epithelial cells transformed by the c-Ha-ras oncogene.

作者信息

Calaf G M, Alvarado M E, Hei T K

机构信息

Center for Radiological Research, Columbia University Medical Center, New York, NY, USA.

出版信息

Oncol Rep. 2005 Oct;14(4):885-93. doi: 10.3892/or.14.4.885.

Abstract

Activated oncogenes have been detected in a variety of malignant tumors and altered expressions of certain genes are known to play a functional role in the cancer process. The chemical carcinogen, BP, and the insertion of c-Ha-ras, induced characteristics of transformed phenotypes in a suitable human breast epithelial cell line. Carcinogen-treated and Ha-ras-transfected cells showed a progression of changes in the morphology, anchorage independent growth, invasiveness and tumorigenicity in SCID mice. Tumor growth occurs after a series of molecular events that parallel morphological changes. The aim of this work was to determine the neoplastic phenotypes following treatment with benzo(a)pyrene (BP) and transfection with c-Ha-ras oncogene changes and PCNA, Neu, ErbB-3 and Cytokeratin 18 protein expression in MCF-10F cells, a spontaneously immortalized human breast epithelial cell line. Protein expression was determined by immunofluorescent staining coupled with confocal microscopy. An increased oncoprotein expression in comparison to MCF-10F cells was observed in PCNA, Neu, ErbB-3 and Cytokeratin 18 protein expression in breast epithelial cells transformed with a chemical carcinogen and/or oncogene transfected that are not present in the MCF-10F. This in vitro cancer model can be used as a valuable model in the study of breast carcinogenesis.

摘要

在多种恶性肿瘤中已检测到激活的癌基因,并且已知某些基因的表达改变在癌症进程中发挥功能性作用。化学致癌物苯并芘(BP)以及c-Ha-ras的插入,在合适的人乳腺上皮细胞系中诱导了转化表型的特征。经致癌物处理和Ha-ras转染的细胞在SCID小鼠中显示出形态、锚定非依赖性生长、侵袭性和致瘤性的一系列变化。肿瘤生长发生在一系列与形态变化平行的分子事件之后。这项工作的目的是确定用苯并(a)芘(BP)处理以及用c-Ha-ras癌基因转染后,自发永生化的人乳腺上皮细胞系MCF-10F细胞中的肿瘤表型变化以及PCNA、Neu、ErbB-3和细胞角蛋白18蛋白的表达。通过免疫荧光染色结合共聚焦显微镜确定蛋白表达。在经化学致癌物处理和/或癌基因转染的乳腺上皮细胞中,观察到与MCF-10F细胞相比,PCNA、Neu、ErbB-3和细胞角蛋白18蛋白表达中癌蛋白表达增加,而MCF-10F细胞中不存在这些情况。这种体外癌症模型可作为研究乳腺癌发生的有价值模型。

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