Diagnosis Value of Detection K-Ras Gene Mutations in Patients with Gastric Cancer

Abstract:

Mutations of K-ras gene have been proven to play important roles in human tumor progression. Point mutations of K-ras gene located in codons 61 are gene hot spot mutation regions in patients with gastric cancer. A simple and effective capillary electrophoresis (CE) method for point mutation detection in codon 61 of K-ras gene was established. The polymerase chain reaction (PCR) fragments included codon 61 of K-ras gene were analyzed by single-strand conformation polymorphism (SSCP)-CE method. The CE analysis was performed by using a 1×Tris–borate–EDTA (TBE) buffer containing 3.0% (w/v) poly (ethylene oxide) (PEO) (MW 300 000) under reverse polarity with 15 oC and voltage of 15 kV. Forty-two gastric cancer patients were analyzed using this developed method. K-ras gene mutation was found in 17 cases (40.5 %) and the results showed that K-ras gene mutation in gastric cancer tissue is a usual event, and the SSCP-CE was feasible for mutation detection of K-ras gene in populations. Detection of gene mutation of gastric cancer tissue by PCR - SSCP has a certain value to diagnosis of gastric cancer and to prevent gastric cancer associated with clinical and pathological features, living habits of different patients by detection mutation of K-ras gene codon 61.

• Periodical: Advanced Materials Research (Volumes 396-398)
• Pages: 1470-1474
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.396-398.1470
• Online since: November 2011
• Authors: Ai Mei Zhang, Rong Wang, Kun Sun, Zheng Ping Jia, Hua Xie
• Keywords: Capillary Electrophoresis, Gene Mutation, Human Gastric Cancer, K-Ras, Single Strand Conformation Polymorphism

References

[1] G.C. Burmer, P.S. Rabinovitch and L.A. Loeb, Cancer Res. Vol. 49 (1989), p.2141

[2] D. DeFeo-Jones, E.M. Scolnick, R. Koller and R .Dhar, Nature. Vol. 306 (1983), p.707

[3] K.R. Mitchelson. Methods Mol. Biol. Vol. 162 (2001), p.3

[4] I.G. Got. Hum. Mutat. Vol. 17 (2002), p.475

[5] M. Hashimoto, F. Barany, F. Xu and S.A. Soper, Analyst. Vol. 132 (2007), p.913

[6] M. Orita, H. Iwahama, H. Kanazawa, K. Hayashi and T. Sekiya. Proc. Natl. Acad. Sci. USA. Vol.86 (1989), p.2766

[7] M. Orita, T. Suzuky, T. Sekiya and K. Hayashi. Genomics. Vol.5(1989), p.874

[8] A.S. Cohen, D.Najarian, J.A. Smith and B.L. Karger. J. Chromatogr. Vol.458 (1988), p.323

[9] N. Nataraj, I. Olivos-Glander, N. Kusukawa and Jr.W. Edward Highsmith. Electrophoresis. Vol. 20 (1999), p.1177

DOI: 10.1002/(sici)1522-2683(19990101)20:6<1177::aid-elps1177>3.0.co;2-2

[10] A.K. Bosserhoff, S. Seegers, C. Hellerbrand, J. Scholmerich and R. Buttner. BioTechniques. Vol. 26 (1999), p.1106

[11] J. Ren. J. Chromatogr. B. Vol. 741 (2000), p.115

[12] C. Gelfi, A. Vigano, S. De Palma, S.G. Righetti, E. Corna and F. Zunino. Electrophoresis. Vol. 23 (2002), p.1517

DOI: 10.1002/1522-2683(200205)23:10<1517::aid-elps1517>3.0.co;2-i

[13] D.J. Harrison, A. Manz, Z. Fan and H. Luedi. Anal. Chem. Vol. 64 (1992), p. (1926)

[14] H. Arakawa, K. Vetanala, M. Maeda. J Chromatogr A. Vol. 664 (1994), p.89

[15] I. Kheterpal and R.A. Mathies. Anal. Chem. Vol. 71 (1999), p.31

[16] M.N. Albarghouthi and A.E. Barron. Electrophoresis. Vol. 21 (2000), p.4096

[17] W.Wei and E.S. Yeung. Anal. Chem. Vol. 73 (2001), p.1776

[18] J.Ren. J. Chromatogr. B. Vol. 741 (2000), p.115

[19] I.V. Kourkine, G. Hestekin, N. Buchholz and B.A. Barron. Anal. Chem. Vol. 74 (2002), p.2565

[20] T.A. Felmlee, R.P. Oda and D. A Persing. J. Chromatogr. A. Vol. 717 (1995), p.127

[21] K. Grunewald, J. Lyons and A. Frohlich. Int J Cancer. Vol. 43 (1989), p.1037

[22] T. Kihana, H. Tsuda and T. Hirota. Jpn J Cancer Res. Vol. 80 (1991), p.308

[23] Y. Endo, L. Zhang and R. Katashima. ElectroPhoresis. Vol. 26(17) (2005), p.3380

[24] P.G. Righetti and C. Gelfi. J Chromatogr B. Vol. 697 (1997), p.195