Enhanced anti-cancer effect of a phosphatidylinositol-3 kinase inhibitor and doxorubicin on human breast epithelial cell lines with different p53 and oestrogen receptor status
Yan A. Wang
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Search for more papers by this authorStuart K. Johnson
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Search for more papers by this authorBarry L. Brown
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Search for more papers by this authorLeeza M. McCarragher
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Search for more papers by this authorKaltoom Al-Sakkaf
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Search for more papers by this authorJanice A. Royds
Department of Pathology, University of Otago, Dunedin, New Zealand
Search for more papers by this authorCorresponding Author
Pauline R.M. Dobson
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, Sheffield University Medical School, Beech Hill Road, Sheffield S10 2RX, UKSearch for more papers by this authorYan A. Wang
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Search for more papers by this authorStuart K. Johnson
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Search for more papers by this authorBarry L. Brown
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Search for more papers by this authorLeeza M. McCarragher
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Search for more papers by this authorKaltoom Al-Sakkaf
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Search for more papers by this authorJanice A. Royds
Department of Pathology, University of Otago, Dunedin, New Zealand
Search for more papers by this authorCorresponding Author
Pauline R.M. Dobson
Cell Signalling Group, Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, University of Sheffield Medical School, Beech Hill Road, Sheffield, S10 2RX, United Kingdom
Academic Unit of Endocrinology, Section of Endocrinology and Reproduction, Sheffield University Medical School, Beech Hill Road, Sheffield S10 2RX, UKSearch for more papers by this authorAbstract
New efforts are being focused on signalling pathways as targets for cancer therapy. This particular study was designed to investigate whether blockade of the phosphatidylinositol 3OH-kinase (PI3K) pathway (a survival/anti-apoptosis pathway, overexpressed in various tumours) could sensitise human breast cancer cells to the effect of chemotherapeutics. Doxorubicin (Dox) and LY294002 (LY, a PI3K inhibitor) were used individually or in combination on MDA-MB-231 (p53 mutant, ER-), T47D (p53 mutant, ER+), and MCF-7 (p53 wildtype, ER+) human breast cancer cell lines, and on 184A1, a nonmalignant human breast epithelial cell line (p53 wildtype, ER-). Each drug showed time- and dose-dependent growth inhibition of cell proliferation on all 4 cell lines. The combination of Dox+LY resulted in enhanced cell growth inhibition in MDA-MB-231 and T47D cells, and additive inhibition in MCF-7 and 184A1 cells. Cell cycle analysis showed that Dox+LY enhanced the arrest of MDA-MB-231 and T47D cells in G2 with the appearance of a sub-G1 peak indicating apoptosis/necrosis, a notion supported by enhanced depolarisation of mitochondrial membrane potential in these cell types. The combination also caused a greater additive increase in Cyclin B1. Thus, the synergistic effect of the combination on cell proliferation in some, but not all, breast cancer cells may be through enhanced induction of both G2 arrest and apoptosis, in which p53 may play a role. Substantially lower doses of doxorubicin could be used with low doses of inhibitors of the PI3K pathway, without compromising the anti-cancer effect, but also lowering detrimental side-effects of doxorubicin. This study supports the notion that survival signalling pathways offer special targets for chemotherapy in cancer. © 2008 Wiley-Liss, Inc.
References
- 1 Davidson S. Oncology: principles and practice of medicine, 19th edn. London: Elsevier Science, 2002. 214 p.
- 2 Roymans D,Slegers H. Phosphatidylinositol 3-kinases in tumor progression. Eur J Biochem 2001; 268: 487–98.
- 3 Fry MJ. Phosphoinositide 3-kinase signalling in breast cancer: How big a role might it play? Breast Cancer Res 2001; 3: 304–12.
- 4 Graff JR,Konicek BW,McNulty AM,Wang Z,Houck K,Allen S,Paul JD,Hbaiu A,Goode RG,Sandusky GE,Vessella RL,Neubauer BL. Increased AKT activity contributes to prostate cancer progression by dramatically accelerating prostate tumor growth and diminishing p27Kip1 expression. J Biol Chem 2000; 275: 24500–5.
- 5 Salh B,Marotta A,Wagey R,Sayed M,Pelech S. Dysregulation of phosphatidylinositol 3-kinase and downstream effectors in human breast cancer. Int J Cancer 2002; 98: 148–54.
- 6 Vlahos CJ,Matter WF,Hui KY,Brown RF. A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)- 8-phenyl-4H-1-benzopyran-4-one (LY294002). J Biol Chem 1994; 269: 5241–8.
- 7 Walker EH,Pacold ME,Perisic O,Stephens L,Hawkins PT,Wymann MP,Williams RL. Structural determinants of phosphoinositide 3-kinase inhibition by wortmannin, LY294002, quercetin, myricetin, and staurosporine. Mol Cell 2000; 6: 909–19.
- 8 Meier R,Hemmings BA. Regulation of protein kinase B. J Recept Signal Transduct Res 1999; 19: 121–8.
- 9 Guo K,Nichol R,Skehel P,Dormann D,Weijer CJ,Williams JG,Pears C. A Dictyostelium nuclear phosphatidylinositol phosphate kinase required for developmental gene expression. EMBO J 2001; 20: 6017–27.
- 10 Kim MS,Cheong YP,So HS,Lee KM,Kim TY,Oh J,Chung YT,Son Y,Kim BR,Park R. Protective effects of morphine in peroxynitrite-induced apoptosis of primary rat neonatal astrocytes: potential involvement of G protein and phosphatidylinositol 3-kinase (PI3 kinase). Biochem Pharmacol 2001; 61: 779–86.
- 11 Ruffels J,Griffin M,Dickenson JM. Activation of ERK1/2, JNK and PKB by hydrogen peroxide in human SH-SY5Y neuroblastoma cells: Role of ERK1/2 in H2O2-induced cell death. Eur J Pharmacol 2004; 483: 163–73.
- 12 Nicholson KM,Anderson NG. The protein kinase B/Akt signalling pathway in human malignancy. Cell Signal 2002; 14: 381–95.
- 13 Liu JP,Schlosser R,Ma WY,Dong Z,Feng H,Lui L,Huang XQ,Liu Y,Li DW. Human alphaA- and alphaB-crystallins prevent UVA-induced apoptosis through regulation of PKCalpha, RAF/MEK/ERK and AKT signaling pathways. Exp Eye Res 2004; 79: 393–403.
- 14 Yang Z,Whelan J,Babb R,Bowen BR. An mRNA splice variant of the AFX gene with altered transcriptional activity. J Biol Chem 2002; 277: 8068–75.
- 15 Bouchard C,Marquardt J,Bras A,Medema RH,Eilers M. Myc-induced proliferation and transformation require Akt-mediated phosphorylation of FoxO proteins. EMBO J 2004; 23: 2830–40.
- 16 Orth HM,Kruger KD,Schmoll D,Grempler R,Scherbaum WA,Joost HG,Bornstein SR,Barthel A. Cellular models for the analysis of signaling by protein kinase B and the forkhead transcription factor FKHR (Foxo1a). Regul Pept 2004; 121: 19–24.
- 17 Cantley LC. The phosphoinositide 3-kinase pathway. Science 2002; 296: 1655–7.
- 18 Chang F,Lee JT,Navolanic PM,Steelman LS,Shelton JG,Blalock WL,Franklin RA,McCubrey JA. Involvement of PI3K/Akt pathway in cell cycle progression, apoptosis, and neoplastic transformation: A target for cancer chemotherapy. Leukemia 2003; 17: 590–603.
- 19 Bellacosa A,De Feo D,Godwin AK,Bell DW,Cheng JQ,Altomare DA,Wan M,Dubeau L,Scambia G,Masciullo V,Ferrandina G,Benedetti Panici P, et al. Molecular alterations of the AKT2 oncogene in ovarian and breast carcinomas. Int J Cancer 1995; 64: 280–5.
- 20 Cheng JQ,Ruggeri B,Klein WM,Sonoda G,Altomare DA,Watson DK,Testa JR. Amplification of AKT2 in human pancreatic cells and inhibition of AKT2 expression and tumorigenicity by antisense RNA. Proc Natl Acad Sci USA 1996; 93: 3636–41.
- 21 Altomare DA,Tanno S,De Rienzo A,Klein-Szanto AJ,Tanno S,Skele KL,Hoffman JP,Testa JR. Frequent activation of AKT2 kinase in human pancreatic carcinomas. J Cell Biochem 2003; 88: 470–6.
- 22 Blume-Jensen P,Hunter T. Oncogenic kinase signalling. Nature 2001; 411: 355–65.
- 23 Thomas CY,Chouinard M,Cox M,Parsons S,Stallings-Mann M,Garcia R,Jove R,Wharen R. Spontaneous activation and signaling by overexpressed epidermal growth factor receptors in glioblastoma cells. Int J Cancer 2003; 104: 19–27.
- 24 Price JT,Tiganis T,Agarwal A,Djakiew D,Thompson EW. Epidermal growth factor promotes MDA-MB-231 breast cancer cell migration through a phosphatidylinositol 3′-kinase and phospholipase C-dependent mechanism. Cancer Res 1999; 59: 5475–8.
- 25 Ignatoski KM,Maehama T,Markwart SM,Dixon JE,Livant DL,Ethier SP. ERBB-2 overexpression confers PI 3′ kinase-dependent invasion capacity on human mammary epithelial cells. Br J Cancer 2000; 82: 666–74.
- 26 Arboleda MJ,Lyons JF,Kabbinavar FF,Bray MR,Snow BE,Ayala R,Danino M,Karlan BY,Slamon DJ. Overexpression of AKT2/protein kinase B beta leads to up-regulation of beta1 integrins, increased invasion, and metastasis of human breast and ovarian cancer cells. Cancer Res 2003; 63: 196–206.
- 27 Nicholson KM,Streuli CH,Anderson NG. Autocrine signalling through erbB receptors promotes constitutive activation of protein kinase B/Akt in breast cancer cell lines. Breast Cancer Res Treat 2003; 81: 117–28.
- 28 Jordan NJ,Gee JM,Barrow D,Wakeling AE,Nicholson RI. Increased constitutive activity of PKB/Akt in tamoxifen resistant breast cancer MCF-7 cells. Breast Cancer Res Treat 2004; 87: 167–80.
- 29 Page C,Lin H-J,Jin Y,Castle VP,Nunez G,Huang M,Lin J. Overexpression of Akt/AKT can modulate chemotherapy-induced apoptosis. Anticancer Res 2000; 20: 407–16.
- 30 Gagnon V,Mathieu I,Sexton E,Leblanc K,Asselin E. AKT involvement in cisplatin chemoresistance of human uterine cancer cells. Gynecol Oncol 2004; 94: 785–95.
- 31 Weinstein IB. Cancer. Addiction to oncogenes-the. Achilles heal of cancer. Science 2002; 297: 63–4.
- 32 Hortobagyi GN. Chemotherapy of breast cancer: A historical perspective. Semin Oncol 1997; 24,S17: 1–4.
- 33 Lehman TA,Modali R,Boukamp P,Stanek J,Bennett WP,Welsh JA,Metcalf RA,Stampfer MR,Fusenig N,Rogan FM. p53 mutations in human immortalized epithelial cell lines. Carcinogenesis 1993; 14: 833–9.
- 34 Alami N,Paterson J,Belanger S,Juste S,Grieshaber CK,Leyland-Jones B. Comparative analysis of xanafide cytotoxicity in breast cancer cell lines. Brit J Cancer 2007; 97: 58–64.
- 35 Seo HS,DeNardo DG,Jacquot Y,Laïos I,Vidal DS,Zambrana CR,Leclercq G,Brown PH. Stimulatory effect of genistein and apigenin on the growth of breast cancer cells correlates with their ability to activate ER alpha. Breast Cancer Res Treat 2006; 99: 121–34.
- 36 Chou TCTalalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enz Regul 1983; 22: 27–55.
- 37 Mills GB,Lu Y,Fang X,Wang H,Eder A,Mao M,Swaby R,Cheng KW,Stokoe D,Siminovitch K,Jaffe R,Gray J. The role of genetic abnormalities of PTEN and the phosphatidylinositol 3-kinase pathway in breast and ovarian tumorigenesis, prognosis, and therapy. Semin Oncol 2001; 28 ( 5 Suppl 16): 125–41.
- 38 O'Gorman DM,McKenna SL,McGahon AJ,Knox KA,Cotter TG. Sensitisation of H160 human leukaemic cells to cytotoxic drug-induced apoptosis by inhibition of PI3-kinase survival signals. Leukaemia 2000; 14: 602–11.
- 39 Mooney LM,Al-Sakkaf KA,Brown BL,Dobson PRM, Apoptotic mechanisms in T47D and MCF-7 human breast cancer cells. Br J Cancer 2002; 87: 909–17.
Citing Literature
