Expression of epidermal growth factor receptor, polyamine levels, ornithine decarboxylase activity, micronuclei, and transglutaminase I in a 7,12-dimethylbenz(a)anthracene-induced hamster buccal pouch carcinogenesis model.
The expression of epidermal growth factor receptor and transglutaminase type I, polyamine (putrescine, spermidine, and spermine) levels, ornithine decarboxylase activity, and micronuclei occurrence were assessed in the 7,12-dimethylbenz(a)anthracene (DMBA)-induced hamster buccal pouch model to elucidate the role and timing of changes in different growth and differentiation markers during carcinogenesis. DMBA (0.5%) in heavy mineral oil was applied to the right buccal pouch 3 times per wk for up to 16 wk; controls received heavy mineral oil alone. Hamsters were killed after 0, 4, 8, and 16 wk. Frozen tissue was chemically analyzed for polyamine levels and ornithine decarboxylase activity and was also used for immunohistochemical analysis of transglutaminase I. Paraffin-embedded sections were used for epidermal growth factor receptor immunohistochemical determinations and for micronucleated cell assays. Hyperplasia was detected by histological analysis at 4 wk, dysplasia with or without papillomatous changes at 8 wk, and squamous cell carcinoma at 16 wk. Epidermal growth factor receptor was not expressed in the normal buccal epithelial layer, at a moderate level in both the superficial keratin and basal cell layers in hyperplastic epithelium, and at very high levels in both dysplasia and squamous cell carcinoma. Transglutaminase I was expressed at a limited level in normal buccal mucosa, was expressed at a low level in the basal layer of hyperplastic lesions, was somewhat elevated in dysplasia, and was markedly enhanced in squamous cell carcinoma. Putrescine and spermidine levels and ornithine decarboxylase activity increased dramatically after 8 and 16 wk of DMBA. Micronucleated cells increased after 4 wk of DMBA treatment, that high level sustained during all stages of carcinogenesis. We suggest that these biological markers could be excellent intermediate end points in assessing the effects of various chemopreventive agents to be tested in the hamster buccal pouch model and in human clinical trials.
Shin
D M
DM
Department of Medical Oncology, M.D. Anderson Cancer Center, Houston, TX 77030.
Gimenez
I B
IB
Lee
J S
JS
Nishioka
K
K
Wargovich
M J
MJ
Thacher
S
S
Lotan
R
R
Slaga
T J
TJ
Hong
W K
WK
D015127:9,10-Dimethyl-1,2-benzanthracene; D000818:Animals; D000911:Antibodies, Monoclonal; D006224:Cricetinae; D066246:ErbB Receptors / Q000378:metabolism*; D006651:Histocytochemistry; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000378:metabolism*; D008647:Mesocricetus; D048629:Micronuclei, Chromosome-Defective / Q000648:ultrastructure*; D009062:Mouth Neoplasms / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D009955:Ornithine Decarboxylase / Q000378:metabolism*; D011073:Polyamines / Q000378:metabolism*; D013997:Time Factors; D011503:Transglutaminases / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D011073:Polyamines; D015127:9,10-Dimethyl-1,2-benzanthracene; D011503:Transglutaminases; D066246:ErbB Receptors; D009955:Ornithine Decarboxylase