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Mutational analysis of HRAS and KRAS genes in oral carcinoma cell lines

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Abstract

RAS overexpression and its active mutations are involved in malignant tumorigenesis. However, the mutation rates in oral carcinoma cells differ between populations. In the present study, genomic DNA of oral carcinoma cells (HOC313, TSU, HSC2, HSC3, KOSC2, KOSC3, SCCKN, OSC19, Ca9.22, and Ho1u1 cells) or normal gingival fibroblasts (GF12 cells) derived from a Japanese population were amplified by polymerase chain reaction using primer sets, spanning HRAS and KRAS exons. Nucleotide substitutions were analyzed by single strand conformation polymorphism. In contrast to no substitutions in KRAS, nine different substitutions were detected in HRAS. Of the nine, six substitutions were located at intron 1 (HSC2 and HSC3 cells) or intron 2 (HSC3, SCCKN and Ca9.22 cells), and one each of exon 1 (all cells), exon 2 (HOC313, TSU, HSC2 and HSC3 cells) and the 5′ upstream region (all cells). Substitutions at exons 1 and 2 did not affect the amino acid sequence; the exon 1 substitution was positioned at the 5′ untranslated region, which may be a single nucleotide polymorphism (SNP) sequence because all the cells were isolated from a Japanese population, and the mutations at exon 2 was a silent mutation. A substitution at the 5′ upstream region was an SNP. These data demonstrate that SNPs and point mutations observed in HRAS do not change the amino acid sequence, and suggest that the mutations affecting the amino acid sequence may be a rare event in oral carcinomas of the Japanese population.

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Acknowledgments

We would like to thank Dr. T. Sato (The Nippon Dental University College at Tokyo) for a generous gift of GF12 cells and Dr. J. D’Armiento (Columbia University) for critically reading the manuscript. This study was performed under the curriculum of Research for Life Dental Sciences at School of Life Dentistry at Tokyo, The Nippon Dental University.

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Authors and Affiliations

  1. Department of Biochemistry, The Nippon Dental University, School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan

    Sachiko Maemoto, Megumi Yumoto, Masato Ibata, Sho Torizuka, Naohumi Ozawa, Shunsuke Tatsumi, Moeko Hashido, Masako Morikawa, Genta Maeda & Kazushi Imai

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  1. Sachiko Maemoto
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  2. Megumi Yumoto
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  3. Masato Ibata
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  4. Sho Torizuka
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  5. Naohumi Ozawa
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  6. Shunsuke Tatsumi
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  7. Moeko Hashido
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  8. Masako Morikawa
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  9. Genta Maeda
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  10. Kazushi Imai
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Correspondence to Kazushi Imai.

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S. Maemoto, M. Yumoto, M. Ibata, S. Torizuka, N. Ozawa, S. Tatsumi and M. Hashido contributed equally to this work.

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Maemoto, S., Yumoto, M., Ibata, M. et al. Mutational analysis of HRAS and KRAS genes in oral carcinoma cell lines. Odontology 100, 149–155 (2012). https://doi.org/10.1007/s10266-011-0032-3

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  • DOI: https://doi.org/10.1007/s10266-011-0032-3

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