Abstract
Abstract: Previously, we identified a novel correlation between the upregulated expression of telomerase (hTERT) and cytochrome P450 1A1 (CYP1A1) in A549 human lung cancer cell line. The expression correlation was confirmed by silencing CYP1A1 expression using siRNA technology and observing a silencing of hTERT transcription. Furthermore, silencing CYP1A1 and subsequently downregulating hTERT resulted in the reduction of cancer cell viability by more than 40%, which appeared as early as 24 hours after the treatment. The concomitant downregulation of CYP1A1 and hTERT resulted in rapid cell death. This finding can be further exploited to develop new molecular targets for the treatment of lung cancer.
Key words: siRNA, gene knockdown, CYP1A1, hTERT, transfection
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Abbreviations used
- AhR
aryl hydrocarbon receptor
- CYP1A1
cytochrome P450 1A1
- hTERT
telomerase
References
- 1.Elbashir S.M., Harborth J., Lendeckel W., Yalcin A., Klaus W., Tuschl T. Duplexes of 21-nucleotide RNAs mediate RNA interference in mammalian cell culture. Nature. 2001;411:494–498. doi: 10.1038/35078107. [DOI] [PubMed] [Google Scholar]
- 2.Hannon G.J., Rossi J.J. Unlocking the potential of the human genome with RNA interference. Nature. 2004;8S:431–371. doi: 10.1038/nature02870. [DOI] [PubMed] [Google Scholar]
- 3.Wilda M., Fuchs U., Wossmann W., Borkhardt A. Killing of leukemic cells with a BCR/ABL fusion gene by RNA interference (RNAi) Oncogene. 2002;37:5716–5724. doi: 10.1038/sj.onc.1205653. [DOI] [PubMed] [Google Scholar]
- 4.Shammas M.A., Koley H., Batchu R.B., Berheau R.C., Protopopov A., Muncshi N.C., Goyal R.J. Telomerase inhibition by siRNA causes senescene and apoptosis in Barretłs adenocarcinoma cells: mechanism and therapeutic potential. Mol. Cancer. 2005;4:1–14. doi: 10.1186/1476-4598-4-24. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5.Flynn M.A., Casey D.G., Todryk S.M., Mohon B.P. Efficient delivery of small interfering RNA for inhibition of IL-12p40 expression in vivo. J. Inflammation. 2004;1:1–12. doi: 10.1186/1476-9255-1-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6.Shervington A., Mohammed K., Patel R., Lea R. Identification of a novel co-transcription of P450/1A1 with telomerase in A549. Gene. 2007;388:100–117. doi: 10.1016/j.gene.2006.10.010. [DOI] [PubMed] [Google Scholar]
- 7.Guigal N., Seree E., Bourgarel-Rey V., Barra Y. Induction of CYP1A1 by serum independent of AhR pathway. Biochem. Biophys. Res. Commun. 2000;267:572–576. doi: 10.1006/bbrc.1999.1959. [DOI] [PubMed] [Google Scholar]
- 8.Iba M.M., Scholl H., Fung J., Thomas P.E., Alan J. Induction of pulmonary CYP1A1 by nicotine. Xenobiotica. 1995;28:827–843. doi: 10.1080/004982598239083. [DOI] [PubMed] [Google Scholar]
- 9.Guengerich F.P. Forging the links between metabolism and carcinogenesis. Mutat. Res. 2001;488:195–209. doi: 10.1016/S1383-5742(01)00059-X. [DOI] [PubMed] [Google Scholar]
- 10.Guengerich F.P. Metabolism of chemical carcinogens. Carcinogenesis. 2000;21:345–351. doi: 10.1093/carcin/21.3.345. [DOI] [PubMed] [Google Scholar]
- 11.Jean-Charles S., Moon C., Wang L., Hittelman W., Jang S., Sun S., Lee J., Liu D., Kurie J., Morice R., Lee J., Hong W., Mao L. Effects of N-(4-Hydroxyphenyl)retinamide on hTERT expression in the bronchial epithelium of cigarette smokers. J. Natl. Cancer Inst. 2001;93:1257–1263. doi: 10.1093/jnci/93.16.1257. [DOI] [PubMed] [Google Scholar]
- 12.Hiyama K., Hiyama E. Detection of telomerase activity in lung cancer tissues. Methods Mol. Med. 2003;74:401–412. doi: 10.1385/1-59259-323-2:401. [DOI] [PubMed] [Google Scholar]
- 13.Xinarianos G. Telomerase activity in non-small cell lung carcinomas correlates with smoking status. Int. J. Oncol. 1999;15:961–965. doi: 10.3892/ijo.15.5.961. [DOI] [PubMed] [Google Scholar]
- 14.Albanell J., Lonardo F., Rusch V., Engelhardt M., Langenfeld J., Han W., Klimstra D., Venkatraman E., Moore M.A., Dmitrovsky E. High telomerase activity in primary lung cancers: association with increased cell proliferation rates and advanced pathologic stage. J. Natl. Cancer Inst. 1997;89:1609–1615. doi: 10.1093/jnci/89.21.1609. [DOI] [PubMed] [Google Scholar]
- 15.Chi J., Chang N.Y., Wang N.N., Chang D.S., Dunphy N., Brown P.O. Genome wide view of gene silencing by small interfering RNAs. Proc. Natl. Acad. Sci. U.S.A. 2003;100:6343–3646. doi: 10.1073/pnas.1037853100. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16.Ambion Inc. siRNA Target Finder. http://www.ambion.com/techlib/misc/siRNA.
- 17.Lekas P., Tin K.L., Lee C., Prokipcak R.D. The human cytochrome P450 1A1 mRNA is rapidly degraded in HepG2 cells. Arch. Biochem. Biophys. 2000;3849:311–318. doi: 10.1006/abbi.2000.2115. [DOI] [PubMed] [Google Scholar]
- 18.Nemoto N., Sakural J. Increase of CYP1A1 mRNA and AHH activity by inhibitors of either protein or RNA synthesis in mouse hepatocytes in primary culture. Carcinogenesis. 1991;12:2115–2121. doi: 10.1093/carcin/12.11.2115. [DOI] [PubMed] [Google Scholar]
- 19.Harley G.B., Futcher A.B., Greidea C.W. Telomerase shorten during aging of human fibroblast. Nature. 1990;345:458–460. doi: 10.1038/345458a0. [DOI] [PubMed] [Google Scholar]
- 20.Shay J.W., Roninson J.B. Hallmarks of senescence in carcinogensis and cancer therapy. Oncogene. 2004;12:2919–2933. doi: 10.1038/sj.onc.1207518. [DOI] [PubMed] [Google Scholar]
- 21.Hukkanen J., Lassila A., Paivarinta K., Valanne S., Sarpo S. Induction and regulation of xenobiotic-metabolizing cytochrome P450s in the human A549 lung adenocarcinoma cell line. Am. J. Respir. Cell Mol. Biol. 2000;22:360–366. doi: 10.1165/ajrcmb.22.3.3845. [DOI] [PubMed] [Google Scholar]
- 22.Liu W., Zhang Y., Shen Y., Jiang J., Miao Z., Ding J. Telomerase inhibition is a specific early event in salvicine-treated human lung adenocarcinoma A549 cells. Biochem. Biophys. Res. Commun. 2004;323:660–667. doi: 10.1016/j.bbrc.2004.08.135. [DOI] [PubMed] [Google Scholar]
- 23.Rossi M.J. Inhibition of telomerase activity by ceramide in A549 human lung adenocarcinoma cells. Mol. Med. J. 2000;2000:1–5. [Google Scholar]
- 24.Pratt W., Toft D. Regulation of signaling protein function and trafficking by the hsp90/hsp70-based chaperone machinery. Exp. Biol. Med. 2004;228:111–133. doi: 10.1177/153537020322800201. [DOI] [PubMed] [Google Scholar]
- 25.Carver L.A., Jackiw V., Bradfield C.A. The 90-kDa heat shock protein is essential for Ah receptor signaling in a yeast expression system. J. Biol. Chem. 1994;269:30109–30112. [PubMed] [Google Scholar]
- 26.Holt S.E., Aisner D.L., Baur J., Tesmer V.M., Dy D. Functional requirement of p23 and Hsp90 in telomerase complexes. Genes Dev. 1999;13:817–826. doi: 10.1101/gad.13.7.817. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 27.Chang S., DePinho R.A. Telomerase extracurricular activities. Proc. Natl. Acad. Sci. U.S.A. 2002;99:12520–12522. doi: 10.1073/pnas.212514699. [DOI] [PMC free article] [PubMed] [Google Scholar]