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ToxSci Advance Access published online on January 23, 2006
Toxicological Sciences, doi:10.1093/toxsci/kfj109
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© The Author 2006. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received September 1, 2005
Accepted January 6, 2006

Carcinogenicity

Characterization of 67 kD Laminin Receptor, A Protein Whose Gene is Over-Expressed on Treatment of Cells with Anti-benzo[a]pyrene -7,8-diol-9,10-epoxide

She-Juan An 1, Jia-Kun Chen 2 *, Hua-jie Chen 3, Wei Chang 3, Yi-Guo Jiang 2, Qing-Yi Wei 3, and Xue-Min Chen 3

1 Department of Occupational and Environmental Health, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan Hubei 430030, China; Lung Cancer Institute, Guangdong Provincial People's Hospital Guangzhou 510080, China
2 Institute for Chemical Carcinogenesis, Guangzhou Medical College, Guangzhou, 510182, China
3 Department of Occupational and Environmental Health, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan Hubei 430030, China

* To whom correspondence should be addressed.
Jia-Kun Chen, E-mail: asjuan{at}126.com


  Abstract

The molecular mechanisms potentially related to tumorigenesis induced by anti-benzo[a]pyrene -7,8-diol-9,10-epoxide (anti-BPDE) were investigated by suppression subtractive hybridization of the human bronchial epithelial cells (16HBE) carcinoma induced by BPDE-transformed 16HBE cells (16HBE-C). The 67 kD laminin receptor gene (67LR1) is one of the screened over-expressed genes in 16HBE-C cells when compared with 16HBE. In order to understand the main functions of 67LR1 gene, we amplified the full length of 67LR1 gene using reverse transcription-polymerase chain reaction (RT-PCR) method. The amplified gene products were inserted into pcDNATM 3.1 Directional TOPO expression vector. We then transfected 16HBE cells with this vector and derived stable transfected 16HBE cells lines containing the 67LR1 gene by using lipofectin and G418 selection protocols. The expression products of transfected genes were analyzed by semi-quantitative RT-PCR. Soft agar growth assay was carried out to identify the malignant features of 67LR1 gene. The stable transfected cell lines can form colonies in soft agar. Further, the transfected cells showed morphological changes compared to the control cells, such as the obvious pseudopods. These data suggest that the 67LR1 gene may be related to malignant transformed induced by the anti-BPDE. The 67LR1 protein may be related to the directionality of cell movement.

Keywords: Anti-BPDE; Laminin Receptor; gene; carcinogenesis; pseudopod.
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