Kras gene mutation analysis in experimentally induced hamster buccal pouch carcinoma

Document Type : Original Article

Abstract

The aim of the present study was directed to screen the oncogenic Kras gene mutation in 7,12-dimethylbenz[a]anthracene (DMBA) induced hamster buccal pouch(s) (HBP(s)) carcinoma. Material and methods: Ten Syrian male hamsters, five weeks old, weighing 80-120g were divided into two group(s) (G(s)). GI (negative control): Right HBP mucosa of 5 animals were painted with liquid paraffin alone three times a week for 18 weeks. GII: (DMBA treated group): Right HBP of 5 animals were painted with 0.5% DMBA in paraffin oil 3 times a week for 18 weeks. After termination of the experiment, the HBPs were excised and bisected with one section fixed in 10% neutral buffered formalin, routinely processed and embedded in paraffin blocks in order to be sectioned and examined histologically. The other sections of the fresh tissues were taken and stored at -80°C for DNA extraction followed by amplification of the targeted sequence using the polymerase chain reaction (PCR) technique. Then, the amplified targeted sequences were sequenced using the chain termination method (Sanger method) with a small concentration of dideoxynucleotides to detect the possible presence of Kras gene mutation. Results: Histological sections, using H&E stain, showed moderately to poorly differentiated SCC. DNA sequencing results revealed that Kras gene mutation was absent in all cases. Conclusion: Oncogenic activation of Kras gene is not important in DMBA induced HBP carcinoma.
 

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