Characterization of 67 kD laminin receptor, a protein whose gene is overexpressed on treatment of cells with anti-benzo[a]pyrene-7,8-diol-9,10-epoxide.
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 overexpressed 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 pcDNA 3.1 Directional TOPO expression vector. We then transfected 16HBE cells with this vector and derived stable transfected 16HBE cell lines containing the 67LR1 gene by using lipofectin and G418 selection protocols. The expression products of transfected genes were analyzed by semiquantitative 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 transformation induced by the anti-BPDE. The 67LR1 protein may be related to the directionality of cell movement.
An
She-Juan
SJ
Department of Occupational and Environmental Health, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan Hubei 430030, China.
Chen
Jia-Kun
JK
Chen
Hua-jie
HJ
Chang
Wei
W
Jiang
Yi-Guo
YG
Wei
Qing-Yi
QY
Chen
Xue-Min
XM
D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000633:toxicity; D000818:Animals; D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000235:genetics*; D002294:Carcinoma, Squamous Cell; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000187:drug effects / Q000235:genetics*; D004847:Epithelial Cells; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D012313:RNA / Q000378:metabolism; D017474:Receptors, Laminin / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D017423:Sequence Analysis, RNA; D014162:Transfection
D014408:Biomarkers, Tumor; D017474:Receptors, Laminin; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D012313:RNA