Abstract
Purpose: Aldehyde dehydrogenases class-1A1 (ALDH1A1) and class-3A1 (ALDH3A1) have been associated with resistance to cyclophosphamide (CP) and its derivatives. We have previously reported the downregulation of these enzymes by all-trans retinoic acid (ATRA). Methods: In this study, we used siRNA duplexes as well as retrovirally expressed siRNA to knockdown one or both enzymes together in A549 lung cancer cell line in order to investigate the role of each one in mediating the resistance and the effect of the addition of ATRA. Results: The results show that significant and specific knockdown of each enzyme can be achieved and that each one contributes similarly to cell resistance to 4-hydroperoxycyclophosphamide (4-HC), an active derivative of CP. Added effects were seen when both enzymes were inhibited. The addition of ATRA also exhibited additional inhibitory effects on ALDH activity and increased 4-HC toxicity when added to single siRNA aimed at one of the enzymes. On the other hand, ATRA had minimal and insignificant additional inhibitory effects on ALDH enzyme activity when added to a combination of siRNAs against both enzymes, but still increased 4-HC toxicity beyond that seen with RNAi-mediated inhibition of both enzymes together. Conclusions: We conclude that both enzymes, ALDH1A1 and ALDH3A1 will need to be blocked in order to achieve the highest sensitivity to 4-HC. Furthermore, ATRA increases 4-HC toxicity even when added to a combination of siRNAs against both enzymes, thus suggesting additional mechanisms by which ATRA can increase drug toxicity.
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References
Vasiliou V, Pappa A, Peterson DR (2000) Role of aldehyde dehydrogenase in endogenous and xenobiotic metabolism. Chem Biol Interact 129:1–19
Sophos NA, Pappa A, Ziegler TL, Vasiliou V (2001) Aldehyde dehydrogenase gene superfamily: the 200 update. Chem Biol Interact 130–132:323–337
Sladek NE (2003) Human aldehyde dehydrogenase: potential pathological, pharmacological, and toxicological impact. J Biochem Mol Toxicol 17:7–23
Hilton J (1984) Role of aldehyde dehydrogenase in cyclophosphamide-resistant L1210 leukemia. Cancer Res 44:5156–5160
Manthey CL, Landkamer GJ, Sladek NE (1990) Identification of the mouse aldehyde dehydrogenases important in aldophosphamide detoxification. Cancer Res 50:4991–5002
Sreerama L, Sladek NE (1993) Identification and characterization of a novel class 3 aldehyde dehydrogenase overexpressed in a human breast adenocarcinoma cell line exhibiting oxazaphosphorine-specific acquired resistance. Biochem Pharmacol 45:2487–2505
von Eitzen U, Meier-Tackmann D, Agarwal DP, Goedde HW (1994) Detoxification of cyclophosphamide by human aldehyde dehydrogenase isozymes. Cancer Lett 76:45–49
Yoshida A, Rzhetsky A, Hsu LC, Chang C (1998) Human aldehyde dehydrogenase gene family. Eur J Biochem 251:549–557
Bunting KD, Lindahl R, Townsend AJ (1994) Oxazaphosphorine-specific resistance in human MCF-7 breast carcinoma cell lines expressing transfected rat class 3 aldehyde dehydrogenase. J Biol Chem 269:23197–23203
Bunting KD, Townsend AJ (1996) De novo expression of transfected human class 1 aldehyde dehydrogenase (ALDH) causes resistance to oxazaphosphorine anti-cancer alkylating agents in hamster V79 cell lines. Elevated class 1 ALDH activity is closely correlated with reduction in DNA interstrand cross-linking and lethality. J Biol Chem 271:11884–11890
Magni M, Shammah S, Schiro R, Mellado W, Dalla-Favera R, Gianni AM (1996) Induction of cyclophosphamide-resistance by aldehyde-dehydrogenase gene transfer. Blood 87:1097–1103
Moreb J, Schweder M, Suresh A, Zucali JR (1996) Overexpression of the human aldehyde dehydrogenase class I results in increased resistance to 4-hydroperoxycyclophosphamide. Cancer Gene Ther 3:24–30
Moreb JS, Schweder M, Gray B, Zucali J, Zori R (1998) In vitro selection for K562 cells with higher retrovirally mediated copy number of aldehyde dehydrogenase class-1 and higher resistance to 4-hydroperoxycyclophosphamide. Hum Gene Ther 9:611–619
Moreb JS, Maccow C, Schweder M, Hecomovich J (2000) Expression of antisense RNA to aldehyde dehydrogenase class-1 sensitizes tumor cells to 4-hydroperoxycyclophosphamide in vitro. J Pharmacol Exp Ther 293:390–396
Moreb JS, Gabr A, Vartikar GR, Gowda S, Zucali JR, Mohuczy D (2005) Retinoic acid down-regulates aldehyde dehydrogenase and increases cytotoxicity of 4-hydroperoxycyclophosphamide and acetaldehyde. J Pharmacol Exp Ther 312:339–345
Zamore PD (2001) RNA interference: listening to the sound of silence. Nat Struct Biol 8:746–750
Bernstein E, Denli AM, Hannon GJ (2001) The rest is silence. RNA 7:1509–1521
Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411:494–498
Tuschl T (2002) Expanding small RNA interference. Nat Biotech 20:446–448
Paul CP, Good PD, Winer I, Engelke DR (2002) Effective expression of small interfering RNA in human cells. Nat Biotech 20:505–508
Tuschl T, Borkhardt A (2002) Small interference RNAs: a revolutionary tool for the analysis of gene function and gene therapy. Mol Interv 2:158–167
Sladek NE, Kollander R, Sreerama L, Kiang DT (2002) Cellular levels of aldehyde dehydrogenases (ALDH1A1 and ALDH3A1) as predictors of therapeutic responses to cyclophosphamide-based chemotherapy of breast cancer: a retrospective study. Rational individualization of oxazaphosphorine-based cancer chemotherapeutic regimens. Cancer Chemother Pharmacol 49:309–321
Szeto W, Jiang W, Tice DA, Rubinfeld B, Hollingshead PG, Fong SE, Dugger DL, Pham T, Yansura DG, Wong TA, Grimaldi JC, Corpuz RT, Singh JS, Frantz GD, Devaux B, Crowley CW, Schwall RH, Eberhard DA, Tastelli L, Polakis P, Pennica D (2001) Overexpression of the retinoic aci-responsive gene stra6 in human cancers and its synergistic induction by Wnt-1 and retinoic acid. Cancer Res 61:4197–4205
Sreerama L, Norman SE (2001) Three different stable human breast adenocarcinoma sublines that overexpress ALDH3A1 and certain other enzymes, apparently as a consequence of constitutively upregulated gene transcription mediated by transactivated EpREs (electrophile responsive elements) present in the 5′-upstream regions of these genes. Chem Biol Interact 130–132:247–260
Sreerama L, Sladek NE (1996) Class I and class 3 aldehyde dehydrogenase levels in the human tumor cell lines currently used by the National Cancer Institute to screen for potentially useful antitumor agents. Adv Exp Med Biol 414:81–94
Choudhary S, Xiao T, Vergara LA, Srivastava S, Nees D, Piatigorsky J, Anusari NH (2005) Role of aldehyde dehydrogenase isozymes in the defense of rat lens and human lens epithelial cells against oxidative stress. Invest Ophthalmol Vis Sci 46:259–267
Haselbeck RJ, Hoffman I, Duester G (1999) Distinct functions for Aldh1 and Raldh2 in the control of ligand production for embryonic retinoid signaling pathways. Dev Genet 25:353–364
Pappa A, Brown D, Koutalos Y, DeGregori J, White C, Vasiliou V (2005) Human aldehyde dehydrogenase 3A1 inhibits proliferation and promotes survival of human corneal epithelial cells. J Biol Chem 280: 27998–28006
Formelli F, Cleris L (1993) Synthetic retinoid fenretinide is effective against a human ovarian carcinoma xenograft and potentiates cisplatin activity. Cancer Res 53:5374–5376
Shalinsky DR, Bischoff ED, Gregory ML, Lamph WW, Heyman RA, Hayes JS, Thomazy V, Davies PJ (1996) Enhanced antitumor efficacy of cisplatin in combination with ALRT1057 (9-cis retinoic acid) in human oral squamous carcinoma xenografts in nude mice. Clin Cancer Res 2:511–520
Grunt T, Dittrich E, Offterdinger M, Schneider SM, Dittrich C, Huber H (1998) Effects of retinoic acid and fenretinide on the c-erbB-2 expression, growth and cisplatin sensitivity of breast cancer cells. Br J Cancer 78:79–87
Kalemkerian GP, Ou X (1999) Activity of fenretinide plus chemotherapeutic agents in small-cell lung cancer cell lines. Cancer Chemother Pharmacol 43:145–150
Pettersson F, Colston KW, Dalgleish AG (2001) Retinoic acid enhances the cytotoxic effects of gemcitabine and cisplatin in pancreatic adenocarcinoma cells. Pancreas 23:273–279
Shankar P, Manjunath N, Lieberman J (2005) The prospect of silencing disease using RNA interference. JAMA 293:1367–1373
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Financial support for this work was provided by a grant (to JSM) from the Flight Attendant Medical Research Institute (Miami, FL).
An erratum to this article can be found at http://dx.doi.org/10.1007/s00280-006-0268-8
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Moreb, J.S., Muhoczy, D., Ostmark, B. et al. RNAi-mediated knockdown of aldehyde dehydrogenase class-1A1 and class-3A1 is specific and reveals that each contributes equally to the resistance against 4-hydroperoxycyclophosphamide. Cancer Chemother Pharmacol 59, 127–136 (2007). https://doi.org/10.1007/s00280-006-0233-6
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DOI: https://doi.org/10.1007/s00280-006-0233-6