Specifically Deoxyribozyme of the PTPRO Gene as a Potential Gene Therapy Means for Human Hepatocellular Carcinoma

Abstract:

Protein tyrosine phosphatase receptor-type O (PTPRO) has been described in several forms of cancer as a new member of the PTP family. The tumor suppressor function of PTPRO was evaluated by design and synthesis the 10-23 deoxyribozyme (DRz), thio-modified DRz (DRz-s) and antisense oligonucleotide (asON) of the PTPRO genomic mRNA to detect the catalytic cleavage activity. Firstly, the cDNA fragment of PTPRO gene was amplified from total cellular RNA of the HepG2.2.15 cells by reverse transcription PCR (RT-PCR). Subsequently, the fragments were cloned to pcDNA3.1(+) plasmids and generated a recombinant plasmids, then sifted the positive recombinant plasmids out to amplify. The expression vector of PTPRO mRNA was obtained in vitro transcription by using T7 RNA polymerase. The results of transfection indicated that when PTPRO mRNA gamyed with deoxyribozyme which activity enhanced, so DRz-s were detected with more intensive specific catalytic cleavage activity than DRz by cells transfecting. And the asON wasn't detected with the property.

• Periodical: Advanced Materials Research (Volumes 781-784)
• Pages: 1203-1208
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.781-784.1203
• Online since: September 2013
• Authors: Chun Nian Su, Min Yu
• Keywords: Deoxyribozyme, Gene Therapy, HepG2.2.15, PTPRO, RT-PCR

References

[1] R.G. Fahmy, A. Waldman, G. Zhang, A. Mitchell, N., Tedla, H., Cai C.R., Geczy, C.N., Chesterman, M., Perry, L.M. Khachigian, J. Nat. Biotechnol. Vol. 24(2006), p.856.

DOI: 10.1038/nbt1225

[2] L. Yen, S.M. Strittmatter, R.G. Kalb, Ann. Neurol. Vol. 46(1999), p.366.

[3] S. Chakraborti, A.C. Banerjea, Molec. Ther. Vol. 7(2003), p.817.

[4] Z.X. Lu, Ye, M., Yan, G.R., Li, Q., Tang, M., Lee, L.M., Sun, L.Q., Y. Cao, Cancer Gene Ther. Vol. 12(2005), p.647.

[5] H. Takahashi, Hamazaki, H., Habu, Y., Hayashi, M., Abe, T., Miyano-Kurosaki, N., H. Takaku, FEBS Lett. P. Vol. 560(2004), p.69.

DOI: 10.1016/s0014-5793(04)00073-0

[6] C.A. Zhao, Zhao, X.D., Yu, H.G., Wu, Y.P., X.Q. Yang, 2003. Zhonghua Er. Ke. Za Zhi. Vol. 41(2003), p.594.

[7] S.T. Crooke, Annu. Rev. Med. Vol. 55(2004), p.61.

[8] S. Schubert, Furste, J.P., Werk, D., Grunert, H.P., Zeichhardt, H., Erdmann, V.A., J. Kurreck, J. Mol. Biol. 339(2004), Vol. p.355–363.

DOI: 10.1016/j.jmb.2004.03.064

[9] R.M. Smith, G.Y. Wu, J. Viral Hepatitis Vol. 10(2003), p.405.

[10] E.M. Westerhout, Ooms, M., Vink, M., Das, A.T., B. Berkhout, Nucleic Acids Res. Vol. 33(2005), p.796.

[11] J.A. Wilson, C.D. Richardson, J. Virol. Vol. 79(2005), p.7050.

[12] Y. Li and R.R. Breaker Curr Opin Struct Biol, Vol. 9(1999), p.315.

[13] Y. Li, R.R. Breaker, Proc Natl Acad Sci USA，Vol. 96-6(1999), p.2746.

[14] Y. Liu, R.R. Breaker, Biochemistry, Vol. 39(2000), p.3106.

[15] N. Carmi, R. R, Breaker, Vol. 9(2001), p.2589.

[16] R.R. Breaker, Science, Vol. 290(2000), p. (2095).

[17] R.R. Breaker, Natl Biotechnol, Vol. 15(1997): p.427.

[18] J. Alonso, J. Sasin, N. Bottini, et al. Cell, Vol. 117(2004): p.699.

[19] J.N. Andersen, P.G. Jansen, S.M. Echwald, et al, FASEB, Vol. 18(2004), p.8.

[20] T. Mustelin, G. S Feng, N. Bottini, Biosci, Vol. 7(2002), pp.85-142.

[21] R.C. Aguiar, Y. Yakushijin, S. Kharbanda, et al. Vol. Blood. 94(1999), p.2403.

[22] S. Avraham, R. London, G.A. Tulloch. Gene, Vol. 204 (1997), p.5.

[23] T. Motiwala, H. Kutay, K. Ghoshal, Oncogene, Vol. 22(2003), p.6319.

[24] T. Motiwala, H. Kutay,M. Ghoshal, Proc Natl Acad Sci USA, Vol. 101(2004), p.13844.

[25] T. Mustelin, R. T Abraham, C.E. Rudd. Front. Biosci, Vol. 7(2002), p.918.

[26] Y.J. You Y.P. Chen X.X. Zheng, H. Meltzer,H. Zhang, CancerLetters, Vol. 315(2012), p.138.

[27] T. Motiwala, H. Kutay, K. Ghoshal, Jacob.S. T, Proc. Natl. Acad. Sci. Vol. 101 (2004) p.13844.

[28] R.C. Aguiar, Y. Y Yakushijin, S Kharbanda, M.A. Shipp, Blood, Vol. 94 (1999) , p.2403.

[29] T. Motiwala, S. Majumder, K. Ghoshal, S. T Jacob, J. Biol. Chem. Vol. 284 (2009) , p.455–464.

[30] P.E. Thomas B.L. Wharram, M. Goyal, J.E. Wiggins, L. B Holzman, R.C. Wiggins, J. Biol. Chem. Vol. 269 (1994)p.19953.

DOI: 10.1016/s0021-9258(17)32113-0

[31] T. Motiwala,K. Ghoshal,S. Majumder, S.T. Jacob,C. Plass, Oncogene, Vol. 22, (2003) , p.6319.

[32] T. Motiwala, S. Majumder, H. Kutay, S.T. Jacob, Clin. Cancer Res. Vol. 13(2007) , p.3174.

[33] B. Ramaswamy, S. Majumder, K. Ghoshal, C.L. Shapiro, S.T. Jacob, Mol. Endocrinol. Vol. 23 (2009) p.176.