Rapid Detection of H-RAS Point Mutation Using Two-Step Polymerase Chain Reaction-Restriction Fragment Length Polymorphism

  • Park, Young-Suk (Department of Molecular Genetics, Seoul Medical Science Institute (SeL)) ;
  • Lee, Kyung-Ok (Department of Molecular Genetics, Seoul Medical Science Institute (SeL)) ;
  • Chai, Young-Gyu (Department of Biochemistry, Hanyang University)
  • 투고 : 1996.05.15
  • 발행 : 1996.09.30

초록

Mutations in codon 12, 13 and 61 of one of the three ras genes, H-ras, K-ras and N-ras, convert these genes into active oncogenes. The presence of H-ras gene mutations have important prognostic implications in various cancers. In this study, the H-ras gene mutations were investigated by two-step PCRRFLP in patients with bladder and stomach cancer. For the control experiments, T24 and SK2 cell lines were used. In a total of 36 bladder cancer patient cases, five (13.9%) mutations were found by this method. Of these, point 12 mutations were two (5.6%) cases and point 61 mutations were three (8.3%) cases. On the other hand, H-ras mutation was not found in 29 cases of stomach cancer. The results of the mutated H-ras gene confirmed by direct sequencing analysis were correlated well with PCR analysis. From the sensitivity test, the H-ras mutation was found to have about 0.2% of mutated DNA mingled in normal DNA. In conclusion, the H-ras mutation has a higher clinical Significance in bladder cancer than stomach cancer. Moreover the two-step PCR-RFLP method is sensitive, rapid and relatively simple for clinical work in detecting H-ras point mutations.

키워드

참고문헌

  1. Nucleic Acids Res. v.12 Bos, J.L.;Verlaan-de, V.;Jansem, M.;Veeneman, A.M.;Van Boom, G.H. https://doi.org/10.1093/nar/12.23.9155
  2. Cancer Res. v.49 Bos, J.L.
  3. Br. J. Urol. v.73 Burchill, S.A.;Neal, D.E.;Lunec, J. https://doi.org/10.1111/j.1464-410X.1994.tb07636.x
  4. Cancer Res. v.54 Cerutti, P.;Hussain, P.;Pourzand, C.;Aguilar, F.
  5. Anal. Biochem. v.128 Elder, J.K.;Southern, E.M. https://doi.org/10.1016/0003-2697(83)90369-X
  6. J. Urol. v.117 Evans, D.R.;Irwin, R.J.;Havre, P.A.;Bouchard, J.G.;Prout, G.R. https://doi.org/10.1016/S0022-5347(17)58598-5
  7. Anal. Biochem. v.37 Feinberg, A.D.;Volgelstein, B.A.
  8. Nature v.309 Fujita, J.;Yoshida, O.;Yuasa, Y.;Rhim, J.S.;Hatanaka, M.;Aaronson, S.A. https://doi.org/10.1038/309464a0
  9. Oncogene v.4 Jiang, W.;Kahn, S.M.;Guillem, J.G.;Weinstein, I.B.
  10. J. Korean Med. Sci. v.7 Koh, E.H.;Chung, H.C.;Lee, K.B. https://doi.org/10.3346/jkms.1992.7.2.110
  11. J. Clin. Pathol. v.46 Korn, S.H.;Moerk, P.T.M.;de Goeij, A.F. https://doi.org/10.1136/jcp.46.7.621
  12. Anal. Biochem. v.102 Laborca, C.;Paigen, K. https://doi.org/10.1016/0003-2697(80)90165-7
  13. Anal. Biochem. v.199 Manam, S.;Nicols, W.W. https://doi.org/10.1016/0003-2697(91)90276-Y
  14. Proc. Natl. Acad. Sci. USA v.82 Mannem, V.;Bekesi, E.;Kung, H.F. https://doi.org/10.1073/pnas.82.2.376
  15. Nature v.310 McGrath, J.P.;Capon, D.J.;Goeddel, D.V.;Levinson, A.D. https://doi.org/10.1038/310644a0
  16. Proc. Natl. Acad. Sci. USA v.84 McMahon, G.;Davis, E.;Wogan, G.N. https://doi.org/10.1073/pnas.84.14.4974
  17. Cancer Res. v.50 Meltzer, S.J.;Mane, S.M.;Wood, P.K.;Resau, J.H.;Newkirk, C.;Terzakis, J.A.;Needlemans, S.W.
  18. Jpn. J. Cancer Res. v.81 Nagata, Y.;Abe, M.;Kobayashi, K.;Saiki, S.;Kotake, T.;Yoshikawa, K.;Ueda, R.;Nakayama, E.;Shiku, H. https://doi.org/10.1111/j.1349-7006.1990.tb02502.x
  19. Gastroenterology v.98 Nanus, D.M.;Kelsen, D.P.;Mentle, I.R. https://doi.org/10.1016/0016-5085(90)90019-W
  20. Cancer Res. v.46 O'Hara, B.M.;Oskarsson, M.;Tainsky, M.A.;Blair, D.G.
  21. Proc. Natl. Acad. Sci. USA v.86 Randall, K.S.;Sean, W.;Corey, H.L.;Henry, A.E. https://doi.org/10.1073/pnas.86.16.6230
  22. Nature v.304 Reddy, E.P.;Reynolds, R.K.;Santos, E.;Barbacid, M.
  23. Keio J. Med. v.41 Saito, S. https://doi.org/10.2302/kjm.41.80
  24. Proc. Natl. Acad. Sci. USA v.83 Sakato, H.;Mori, M.;Taira, M.;Yoshida, T.;Matsukawa, S.;Shimizu, K.;Sekiguchi, M.;Terada, M.;Sugimura, T. https://doi.org/10.1073/pnas.83.11.3997
  25. Proc. Natl. Acad. Sci. USA v.81 Sekiya, T.;Hori, H.;Hirohashi, S.;Nishamura, S.;Sugimura, T. https://doi.org/10.1073/pnas.81.15.4771
  26. Cell v.19 Shih, C.;Weinberg, R.A.
  27. Proc. Natl. Acad. Sci. USA v.83 Shimizu, K.M.;Goldfarb, Y.;Guard, M.;Perucho, Y.;Feramisco, E.
  28. Lab. Inves. v.68 Sugano, Kl;Kyogoku, A.;Fukayama, N.;Ohkura, H.;Shimosato, Y.;Sekiya, T.;Kayashi, K.
  29. Urol. Int. v.55 Uchida, T.;Wada, C.;Ishida, H.;Egawa, S.;Ao, T.;Yokoyama, E.;Koshiba, K. https://doi.org/10.1159/000282753
  30. Oncogene Res. v.3 Visvanathan, K.U.;Pocock, R.D.;Summerhayes, I.C.
  31. Proc. Natl. Acad. Sci. USA v.82 Winter, E.;Yamamoto, F.;Almoguera, C.;Perucho, M. https://doi.org/10.1073/pnas.82.22.7575
  32. Jpn. J. Cancer Res. v.77 Yamada, H.;Sakamoto, H.;Taira, M.;Nishimura, S.;Shimosato, Y.;Terada, M.;Sugimura, T.
  33. Jpn. J. Cancer Res. v.77 Yuasa, Y.;Oto, M.;Sato, C.;Namba, M.