Abstract

Changes in nuclear and nucleolar morphometric parameters were investigated by image analysis procedures in human breast MCF-10F epithelial cells expressing different stages of the tumourigenic progression after benzo[a]pyrene (BP) transformation (BP1, BP1-E, and BP1-E1 cell lines), and additionally transfected with the c-Ha-ras oncogene (BP1-Tras cell line). Nuclear pleomorphism was evident in all the transformed cells. The analysis of different morphometric parameters did not show a clear relationship between specific nuclear and nucleolar changes and the expression of the different stages of the tumourigenesis, with the exception of the nucleolar size, which could be associated to the expression of the tumourigenic phenotype, and a nucleolar area/nuclear area ratio, which discriminated the immortalized, the transformed, and the tumourigenic phenotypes from one another. The nuclear morphometric data established for the BP-transformed cells and for the cells additionally transfected with the c-Ha-ras oncogene were suggestive of complex and distinct morphofunctional mechanisms involving the in vitro transformation of the MCF-10F cells. The nuclear changes found in the BP1-Tras cell line were assumed to be related to the additional effects and/or enhanced genomic instability induced by transfection with the ras oncogene.

Highlights

  • Neoplastic transformation is a multistep process that leads cells to morphological and physiological progressive changes due to the inactivation of tumour suppressor genes and the activation of oncogenes that act at different cellular sites [7,18]

  • Changes in nuclear and nucleolar morphometric parameters have been proposed by several authors as tools for breast cancer diagnosis, grading and prognosis [8,22,23,24]

  • The present results suggest that certain nuclear and nucleolar morphometric changes evaluated by image analysis can be associated to the in vitro neoplastic progression in MCF-10F cells

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Summary

Introduction

Neoplastic transformation is a multistep process that leads cells to morphological and physiological progressive changes due to the inactivation of tumour suppressor genes and the activation of oncogenes that act at different cellular sites [7,18]. L.F. Barbisan et al / Nuclear and nucleolar image analysis of human breast epithelial cells size, shape and chromatin texture have been demonstrated during in vitro neoplastic transformation induced by chemical carcinogens and c-Ha-ras oncogene transfection in murine and human cells [13, 14,16]. Immortalized human breast epithelial cell lines, MCF-10A and F, have been used as experimental models to study the in vitro neoplastic progression induced by chemical carcinogens and/or c-Ha-ras transfection [2,4,5,21]. We investigated by image analysis procedures nuclear and nucleolar morphometric changes in human breast epithelial cell lines transformed by benzo[a]pyrene and transfected with the c-Ha-ras oncogene

Cell lines and culture conditions
Cell preparations and staining
Image analysis
Results
Discussion
Full Text
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