Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
BMC Cancer
Published in: BMC Cancer 1/2015

Open Access 01-12-2015 | Research article

Phosphatidylinositol- 3-kinase inhibitor induces chemosensitivity to a novel derivative of doxorubicin, AD198 chemotherapy in human bladder cancer cells in vitro

Authors: Dmitriy Smolensky, Kusum Rathore, Maria Cekanova

Published in: BMC Cancer | Issue 1/2015

Login to get access

Abstract

Background

Doxorubicin (Dox) is widely used to treat progressed bladder cancer after transurethral resection. The use of Dox-chemotherapy has been limited due to induced drug resistance and cumulative cardiotoxic effects. N-benzyladriamycin-14-valerate (AD198), a novel derivative of Dox, has a potential to become a more effective treatment than Dox by overcoming drug resistance and cardio-toxicity as shown in the rodent model of lymphoma in vivo. The purpose of this study was to compare the efficacy of Dox and AD198 and explore their mechanisms in inhibition on human bladder cancer cells in vitro.

Methods

We evaluated the effects of Dox and AD198 on cell viability of human transitional cell carcinoma (TCC) cell lines T24 and UMUC3 by MTS assay in vitro. The effects of Dox and AD198 on cell apoptosis were determined by caspase 3/7 assay, generation of reactive oxygen species (ROS), and Western Blotting (WB) analysis.

Results

AD198 was more effective than Dox in inhibition of cell viability of T24 and UMUC3 cells in vitro. Both Dox and AD198 significantly increased the generation of ROS and induced apoptosis in caspase-dependent and -independent manner in T24 and UMUC3 cells. AD 198 induced significantly higher production of ROS as compared to Dox in human TCC cells. Dox and AD198 activated the pro-apoptotic p38 MAPK pathway; however, on the other hand also increased phosphorylation of AKT, an anti-apoptotic signaling pathway, in T24 and UMUC3 cells. Combined treatment of PI3K inhibitor (LY294002) with Dox or AD198 inhibited cell viability of T24 and UMUC3 cells more effectively than any of drug treatments alone.

Conclusions

These data suggest that AD198 as novel derivative of Dox, could be a used as effective treatment for bladder cancer. Dox and AD198 induced PI3K/AKT signaling pathway that is a one of the indicators of pro-survival and possible drug-resistance mechanisms of chemotherapies in bladder cancer. Combined therapies of Dox or AD198 with inhibitors of PI3K/AKT signaling pathway might lead to more effective treatment outcome for patients diagnosed with bladder cancer based on our in vitro experiments.
Literature
1.
2.
3.
4.
5.
Freedman ND, Silverman DT, Hollenbeck AR, Schatzkin A, Abnet CC. Association between smoking and risk of bladder cancer among men and women. JAMA. 2011;306(7):737–45.PubMedCentralCrossRefPubMed
6.
Solsona E, Iborra I, Ricos JV, Monros JL, Casanova J, Dumont R. Effectiveness of a single immediate mitomycin C instillation in patients with low risk superficial bladder cancer: short and long-term followup. J Urol. 1999;161(4):1120–3.CrossRefPubMed
7.
Ali-el-Dein B, Nabeeh A, el-Baz M, Shamaa S, Ashamallah A. Single-dose versus multiple instillations of epirubicin as prophylaxis for recurrence after transurethral resection of pTa and pT1 transitional-cell bladder tumours: a prospective, randomized controlled study. Br J Urol. 1997;79(5):731–5.CrossRefPubMed
8.
Lamm DL, Blumenstein BA, Crawford ED, Montie JE, Scardino P, Grossman HB, et al. A randomized trial of intravesical doxorubicin and immunotherapy with bacille Calmette-Guerin for transitional-cell carcinoma of the bladder. N Engl J Med. 1991;325(17):1205–9.CrossRefPubMed
9.
Herr HW, Schwalb DM, Zhang ZF, Sogani PC, Fair WR, Whitmore Jr WF, et al. Intravesical bacillus Calmette-Guerin therapy prevents tumor progression and death from superficial bladder cancer: ten-year follow-up of a prospective randomized trial. J Clin Oncol. 1995;13(6):1404–8.PubMed
10.
11.
Keizer HG, Pinedo HM, Schuurhuis GJ, Joenje H. Doxorubicin (adriamycin): A critical review of free radical-dependent mechanisms of cytotoxicity. Pharmacol Ther. 1990;47(2):219–31.CrossRefPubMed
12.
Bodley A, Liu LF, Israel M, Seshadri R, Koseki Y, Giuliani FC, et al. DNA Topoisomerase II-mediated Interaction of Doxorubicin and Daunorubicin Congeners with DNA. Cancer Res. 1989;49(21):5969–78.PubMed
13.
Wassermann K, Markovits J, Jaxel C, Capranico G, Kohn KW, Pommier Y. Effects of morpholinyl doxorubicins, doxorubicin, and actinomycin D on mammalian DNA topoisomerases I and II. Mol Pharmacol. 1990;38(1):38–45.PubMed
14.
Capranico G, Kohn KW, Pommier Y. Local sequence requirements for DNA cleavage by mammalian topoisomerase II in the presence of doxorubicin. Nucleic Acids Res. 1990;18(22):6611–9.PubMedCentralCrossRefPubMed
15.
Mizutani H, Tada-Oikawa S, Hiraku Y, Kojima M, Kawanishi S. Mechanism of apoptosis induced by doxorubicin through the generation of hydrogen peroxide. Life Sci. 2005;76(13):1439–53.CrossRefPubMed
16.
Wang S, Konorev EA, Kotamraju S, Joseph J, Kalivendi S, Kalyanaraman B. Doxorubicin Induces apoptosis in normal and tumor cells via distinctly different mechanisms: intermediacy of H2O2- and p53-dependent pathways. J Biol Chem. 2004;279(24):25535–43.CrossRefPubMed
17.
Tsang WP, Chau SPY, Kong SK, Fung KP, Kwok TT. Reactive oxygen species mediate doxorubicin induced p53-independent apoptosis. Life Sci. 2003;73(16):2047–58.CrossRefPubMed
18.
Dalton WS, Grogan TM, Meltzer PS, Scheper RJ, Durie BG, Taylor CW, et al. Drug-resistance in multiple myeloma and non-Hodgkin’s lymphoma: detection of P-glycoprotein and potential circumvention by addition of verapamil to chemotherapy. J Clin Oncol. 1989;7(4):415–24.PubMed
19.
20.
Frishman WH, Sung HM, Yee HCM, Liu LL, Einzig AI, Dutcher J, et al. Cardiovascular toxicity with cancer chemotherapy. Curr Probl Cardiol. 1996;21(4):227–86.CrossRef
21.
Lothstein L, Wright HM, Sweatman TW, Israel M. N-benzyladriamycin-14-valerate and drug resistance: correlation of anthracycline structural modification with intracellular accumulation and distribution in multidrug resistant cells. Oncol Res. 1992;4(8–9):341–7.PubMed
22.
Lothstein L, Savranskaya L, Barrett CM, Israel M, Sweatman TW. N-benzyladriamycin-14-valerate (AD 198) activates protein kinase C-delta holoenzyme to trigger mitochondrial depolarization and cytochrome c release independently of permeability transition pore opening and Ca2+ influx. Anticancer Drugs. 2006;17(5):495–502.CrossRefPubMed
23.
Hofmann PA, Israel M, Koseki Y, Laskin J, Gray J, Janik A, et al. N-Benzyladriamycin-14-valerate (AD 198): a non-cardiotoxic anthracycline that is cardioprotective through PKC-ε activation. J Pharmacol Exp Ther. 2007;323(2):658–64.CrossRefPubMed
24.
Edwards SK, Moore CR, Liu Y, Grewal S, Covey LR, Xie P. N-benzyladriamycin-14-valerate (AD 198) exhibits potent anti-tumor activity on TRAF3-deficient mouse B lymphoma and human multiple myeloma. BMC Cancer. 2013;13(1):481.PubMedCentralCrossRefPubMed
25.
Rathore K, Cekanova M. A novel derivative of doxorubicin, AD198, inhibits canine transitional cell carcinoma and osteosarcoma cells in vitro. J Drug Des Dev Ther. 2015;2015(9):5323–35.
26.
Sweatman TW, Seshadri R, Israel M. Pharmacology of N-benzyladriamycin-14-valerate in the rat. Cancer Chemother Pharmacol. 1999;43(5):419–26.CrossRefPubMed
27.
Cai C, Lothstein L, Morrison RR, Hofmann PA. Protection from doxorubicin-induced cardiomyopathy using the modified anthracycline N-benzyladriamycin-14-valerate (AD 198). J Pharmacol Exp Ther. 2010;335(1):223–30.PubMedCentralCrossRefPubMed
28.
Duriez P, Shah GM. Cleavage of poly (ADP-ribose) polymerase: a sensitive parameter to study cell death. Biochem Cell Biol. 1997;75(4):337–49.CrossRefPubMed
29.
Soldani C, Scovassi A. Poly (ADP-ribose) polymerase-1 cleavage during apoptosis: an update. Apoptosis. 2002;7(4):321–8.CrossRefPubMed
30.
Franke TF, Kaplan DR, Cantley LC. PI3K: downstream AKTion blocks apoptosis. Cell. 1997;88(4):435–7.CrossRefPubMed
31.
Wold LE, Aberle NS, Ren J. Doxorubicin induces cardiomyocyte dysfunction via a p38 MAP kinase-dependent oxidative stress mechanism. Cancer Detect Prev. 2005;29(3):294–9.CrossRefPubMed
32.
Deng YT, Huang HC, Lin JK. Rotenone induces apoptosis in MCF‐7 human breast cancer cell‐mediated ROS through JNK and p38 signaling. Mol Carcinog. 2010;49(2):141–51.PubMed
33.
Garnick MB, Schade D, Israel M, Maxwell B, Richie JP. Intravesical doxorubicin for prophylaxis in the management of recurrent superficial bladder carcinoma. J Urol. 1984;131(1):43–6.PubMed
34.
Chen C-H, Yang H-J, Shun C-T, Huang C-Y, Huang K-H, Yu H-J, et al. A cocktail regimen of intravesical mitomycin-C, doxorubicin, and cisplatin (MDP) for non-muscle-invasive bladder cancer. Urol Oncol. 2012;30(4):421–7.CrossRefPubMed
35.
Kimiya K, Naito S, Soejima T, Sakamoto N, Kotoh S, Kumazawa J, et al. Establishment and characterization of doxorubicin-resistant human bladder cancer cell line, KK47/ADM. J Urol. 1992;148(2 Pt 1):441–5.PubMed
36.
Lothstein L, Israel M, Sweatman TW. Anthracycline drug targeting: cytoplasmic versus nuclear–a fork in the road. Drug Resist Updat. 2001;4(3):169–77.CrossRefPubMed
37.
Ganapathi R, Grabowski D, Sweatman TW, Seshadri R, Israel M. N-benzyladriamycin-14-valerate versus progressively doxorubicin-resistant murine tumours: cellular pharmacology and characterisation of cross-resistance in vitro and in vivo. Br J Cancer. 1989;60(6):819–26.PubMedCentralCrossRefPubMed
38.
Lothstein L, Savranskaya L, Sweatman TW. N-Benzyladriamycin-14-valerate (AD 198) cytotoxicty circumvents Bcr-Abl anti-apoptotic signaling in human leukemia cells and also potentiates imatinib cytotoxicity. Leuk Res. 2007;31(8):1085–95.CrossRefPubMed
39.
Harstrick A, Vanhoefer U, Schieucher N, Schroeder J, Baumgart J, Scheulen M, et al. Activity of N-benzyl-adriamycin-14-valerate (AD198), a new anthracycline derivate, in multidrug resistant human ovarian and breast carcinoma cell lines. Anticancer Drugs. 1995;6(5):681–5.CrossRefPubMed
40.
Lothstein L, Wright H, Sweatman T, Israel M. N-benzyladriamycin-14-valerate and drug resistance: correlation of anthracycline structural modification with intracellular accumulation and distribution in multidrug resistant cells. Oncol Res. 1991;4(8–9):341–7.
41.
Gamen S, Anel A, Lasierra P, Alava MA, Martinez-Lorenzo MJ, Piñeiro A, et al. Doxorubicin-induced apoptosis in human T-cell leukemia is mediated by caspase-3 activation in a Fas-independent way. FEBS Lett. 1997;417(3):360–4.CrossRefPubMed
42.
Bernuzzi F, Recalcati S, Alberghini A, Cairo G. Reactive oxygen species-independent apoptosis in doxorubicin-treated H9c2 cardiomyocytes: role for heme oxygenase-1 down-modulation. Chem Biol Interact. 2009;177(1):12–20.CrossRefPubMed
43.
McCubrey JA, Steelman LS, Abrams SL, Lee JT, Chang F, Bertrand FE, et al. Roles of the RAF/MEK/ERK and PI3K/PTEN/AKT pathways in malignant transformation and drug resistance. Adv Enzyme Regul. 2006;46(1):249–79.CrossRefPubMed
44.
Misra S, Ghatak S, Toole BP. Regulation of MDR1 expression and drug resistance by a positive feedback loop involving hyaluronan, phosphoinositide 3-kinase, and ErbB2. J Biol Chem. 2005;280(21):20310–5.CrossRefPubMed
45.
Bezler M, Hengstler JG, Ullrich A. Inhibition of doxorubicin-induced HER3-PI3K-AKT signalling enhances apoptosis of ovarian cancer cells. Mol Oncol. 2012;6(5):516–29.CrossRefPubMed
46.
Ghebeh H, Lehe C, Barhoush E, Al-Romaih K, Tulbah A, Al-Alwan M, et al. Research article Doxorubicin downregulates cell surface B7-H1 expression and upregulates its nuclear expression in breast cancer cells: role of B7-H1 as an anti-apoptotic molecule. 2010.
47.
Lai J-P, Sandhu DS, Moser CD, Cazanave SC, Oseini AM, Shire AM, et al. Additive effect of apicidin and doxorubicin in sulfatase 1 expressing hepatocellular carcinoma in vitro and in vivo. J Hepatol. 2009;50(6):1112–21.PubMedCentralCrossRefPubMed
48.
Sun CH, Chang YH, Pan CC. Activation of the PI3K/Akt/mTOR pathway correlates with tumour progression and reduced survival in patients with urothelial carcinoma of the urinary bladder. Histopathology. 2011;58(7):1054–63.CrossRefPubMed
49.
Network TCGAR. Comprehensive molecular characterization of urothelial bladder carcinoma. Nature. 2014;507(7492):315–22.CrossRef
50.
Knowles MA, Platt FM, Ross RL, Hurst CD. Phosphatidylinositol 3-kinase (PI3K) pathway activation in bladder cancer. Cancer Metastasis Rev. 2009;28(3–4):305–16.PubMedCentralCrossRefPubMed
51.
Seront E, Pinto A, Bouzin C, Bertrand L, Machiels J-P, Feron O. PTEN deficiency is associated with reduced sensitivity to mTOR inhibitor in human bladder cancer through the unhampered feedback loop driving PI3K/Akt activation. Br J Cancer. 2013;109(6):1586–92.PubMedCentralCrossRefPubMed
52.
Martin-Doyle W, Kwiatkowski DJ. Molecular biology of bladder cancer. Hematol Oncol Clin North Am. 2015;29(2):191–203.CrossRefPubMed
53.
Baldo P, Cecco S, Giacomin E, Lazzarini R, Ros B, Marastoni S. mTOR pathway and mTOR inhibitors as agents for cancer therapy. Curr Cancer Drug Targets. 2008;8(8):647–65.CrossRefPubMed
54.
Wu D, Tao J, Xu B, Qing W, Li P, Lu Q, et al. Phosphatidylinositol 3-Kinase inhibitor LY294002 suppresses proliferation and sensitizes doxorubicin chemotherapy in bladder cancer cells. Urol Int. 2011;87(1):105–13.CrossRefPubMed
55.
Saal LH, Johansson P, Holm K, Gruvberger-Saal SK, She Q-B, Maurer M, et al. Poor prognosis in carcinoma is associated with a gene expression signature of aberrant PTEN tumor suppressor pathway activity. Proc Natl Acad Sci. 2007;104(18):7564–9.PubMedCentralCrossRefPubMed
56.
Tanaka M, Grossman HB. In vivo gene therapy of human bladder cancer with PTEN suppresses tumor growth, downregulates phosphorylated Akt, and increases sensitivity to doxorubicin. Gene Ther. 2003;10(19):1636–42.CrossRefPubMed
57.
Wang DS, Rieger‐Christ K, Latini JM, Moinzadeh A, Stoffel J, Pezza JA, et al. Molecular analysis of PTEN and MXI1 in primary bladder carcinoma. Int J Cancer. 2000;88(4):620–5.CrossRefPubMed
Metadata
Title
Phosphatidylinositol- 3-kinase inhibitor induces chemosensitivity to a novel derivative of doxorubicin, AD198 chemotherapy in human bladder cancer cells in vitro
Authors
Dmitriy Smolensky
Kusum Rathore
Maria Cekanova
Publication date
01-12-2015
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2015
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/s12885-015-1930-5

Other articles of this Issue 1/2015

BMC Cancer 1/2015 Go to the issue

Research article

The impact of supplementary narrative-based information on colorectal cancer screening beliefs and intention

Research article

Under-treatment of elderly patients with ovarian cancer: a population based study

Research article

Combined treatment of Nimotuzumab and rapamycin is effective against temozolomide-resistant human gliomas regardless of the EGFR mutation status

Research article

Methylation status of COX-2 in blood leukocyte DNA and risk of gastric cancer in a high-risk Chinese population

Research article

Human non-small cell lung cancer expresses putative cancer stem cell markers and exhibits the transcriptomic profile of multipotent cells

Research article

Heparan sulfate proteoglycans undergo differential expression alterations in right sided colorectal cancer, depending on their metastatic character
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits