Overcoming chemoresistance in osteosarcoma: the role of autophagy in cell death or survival following chemotherapy

Conference item


Rosa, G., Burczynska, B., Stordal, B. and Roberts, H. 2018. Overcoming chemoresistance in osteosarcoma: the role of autophagy in cell death or survival following chemotherapy. Research Degrees Students' Summer Conference. Middlesex University, London, UK 04 Sep 2018
TitleOvercoming chemoresistance in osteosarcoma: the role of autophagy in cell death or survival following chemotherapy
AuthorsRosa, G., Burczynska, B., Stordal, B. and Roberts, H.
Abstract

Background: For the past 30 years, long-term survival rates in metastatic and recurrent osteosarcoma (OS) patients has remained unchanged. Drug resistance is thought to be the main cause and overcoming this phenomenon is a key step towards greater efficacy with OS therapy. Increasing evidence shows that autophagy, a 'self-degradation' pathway, can act as a protective mechanism to help cancer cells thrive under chemotherapeutic stress. The current study investigated the function of autophagy in two OS cell lines of varying metastatic capacity in response to two standard chemotherapy treatments, doxorubicin and cisplatin. Method: HOS-143B (highly metastatic) and MG-63 (less metastatic) cell lines were treated with doxorubicin (Dox) and cisplatin (Cis) for 48h (1). An acid phosphatase-based proliferation assay was used to calculate IC50s of each drug. Autophagy activation was determined based on the protein expression of p62 and autophagic flux (LC3-I/LC3-II protein conversion), analysed with Western blotting. Gene expression of MAPLC3B and p62/SQSTM1 was measured using RT-PCR. LC3-II punctate formation was detected using immunofluorescence. Results: Both drugs significantly induced autophagic flux in MG63 and HOS-143B cells as shown by increased LC3-I to LC3-II conversion, with Dox increasing LC3-II expression to a higher extent compared with Cis. Immunofluorescence of LC3-II punctate confirmed this pattern. Dox treatment was also associated with reduced p62/SQSTM1 protein, indicating greater autophagic clearance. MAPLC3B and p62/SQSTM1 gene expression was elevated in both Dox treated cell lines. Conclusion: In both highly metastatic HOS-143B cells and less metastatic MG63 cell line, dox treatment induces autophagy to a greater extent when compared with cis. It is envisaged that enhancing our knowledge on the role of autophagy in chemoresistance will help us to identify novel targets for osteosarcoma treatment.

Sustainable Development Goals3 Good health and well-being
Middlesex University ThemeHealth & Wellbeing
Research GroupBiomarkers for Cancer group
ConferenceResearch Degrees Students' Summer Conference
Publication dates
Print04 Sep 2018
Publication process dates
Deposited25 Jul 2022
Accepted04 Aug 2018
Output statusPublished
LanguageEnglish
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Pirmadjid, N., Torrance, H., Owen, H., Alazawi, W. and O'Dwyer, M. 2015. Association between early postoperatively elevated TLR-4 & TLR-5 gene expression and late nosocomial infection in elective high-risk hepato-pancreato-biliary (HPB) surgery. 2015 International Surgical Congress of the Association of Surgeons of Great Britain and Ireland. Manchester, UK 22 - 24 Apr 2015 Oxford University Press (OUP).
T-helper cell polarisation following severe polytrauma
Torrance, H., Brohi, K., Warnes, G., Owen, H., Hinds, C., Pennington, D. and O'Dwyer, M. 2015. T-helper cell polarisation following severe polytrauma. ESICM LIVES 2015: 28th Annual Congress of European Society of Intensive Care Medicine. Berlin, Germany 03 - 07 Oct 2015 Springer Open. https://doi.org/10.1186/2197-425X-3-S1-A848
Post-operative immune suppression is reversible with interferon gamma and independent of IL-6 pathways
Longbottom, E., Torrance, H., Owen, H., Hinds, C., Pearse, R. and O'Dwyer, M. 2015. Post-operative immune suppression is reversible with interferon gamma and independent of IL-6 pathways. ESICM LIVES 2015: 28th Annual Congress of European Society of Intensive Care Medicine. Berlin, Germany 03 - 07 Oct 2015 Springer Open. https://doi.org/10.1186/2197-425X-3-S1-A820
The role of micrornas in the development of hospital acquired infection in polytrauma patients
Owen, H., Torrance, H., Barnes, M., Brohi, K., Knight, J., Hinds, C. and O'Dwyer, M. 2015. The role of micrornas in the development of hospital acquired infection in polytrauma patients. ESICM LIVES 2015: 28th Annual Congress of European Society of Intensive Care Medicine. Berlin, Germany 03 - 07 Oct 2015 Springer Open. https://doi.org/10.1186/2197-425X-3-S1-A35
Changes in gene expression following trauma are related to the age of transfused packed red blood cells
Torrance, H., Vivian, M., Brohi, K., Prowle, J., Pearse, R., Owen, H., Hinds, C. and O'Dwyer, M. 2015. Changes in gene expression following trauma are related to the age of transfused packed red blood cells. Journal of Trauma and Acute Care Surgery. 78 (3), pp. 535-542. https://doi.org/10.1097/TA.0000000000000534
The perioperative immune response
O'Dwyer, M., Owen, H. and Torrance, H. 2015. The perioperative immune response. Current Opinion in Critical Care. 21 (4), pp. 336-342. https://doi.org/10.1097/MCC.0000000000000213
Critical illness-induced bone loss is related to deficient autophagy and histone hypomethylation
Owen, H., Vanhees, I., Gunst, J., Van Cromphaut, S. and Van den Berghe, G. 2015. Critical illness-induced bone loss is related to deficient autophagy and histone hypomethylation. Intensive Care Medicine Experimental. 3. https://doi.org/10.1186/s40635-015-0052-3
Epigenetic regulatory pathways involving microRNAs may modulate the host immune response following major trauma
Owen, H., Torrance, H., Jones, T., Pearse, R., Hinds, C., Brohi, K. and O'Dwyer, M. 2015. Epigenetic regulatory pathways involving microRNAs may modulate the host immune response following major trauma. Journal of Trauma and Acute Care Surgery. 79 (5), pp. 766-772. https://doi.org/10.1097/TA.0000000000000850
Non-invasive and non-destructive measurements of confluence in cultured adherent cell lines
Busschots, S., O'Toole, S., O'Leary, J. and Stordal, B. 2015. Non-invasive and non-destructive measurements of confluence in cultured adherent cell lines. MethodsX. 2, pp. 8-13. https://doi.org/10.1016/j.mex.2014.11.002
Identifying novel hypoxia-associated markers of chemoresistance in ovarian cancer
McEvoy, L., O'Toole, S., Spillane, C., Martin, C., Gallagher, M., Stordal, B., Blackshields, G., Sheils, O. and O'Leary, J. 2015. Identifying novel hypoxia-associated markers of chemoresistance in ovarian cancer. BMC cancer. 15 (1). https://doi.org/10.1186/s12885-015-1539-8
Carboplatin and taxol resistance develops more rapidly in functional BRCA1 compared to dysfunctional BRCA1 ovarian cancer cells
Busschots, S., O'Toole, S., O'Leary, J. and Stordal, B. 2015. Carboplatin and taxol resistance develops more rapidly in functional BRCA1 compared to dysfunctional BRCA1 ovarian cancer cells. Experimental Cell Research. 336 (1), pp. 1-14. https://doi.org/10.1016/j.yexcr.2014.12.001
Humanized culture of periosteal progenitors in allogeneic serum enhances osteogenic differentiation and in vivo bone formation
Roberts, S., Owen, H., Tam, W., Solie, L., Van Cromphaut, S., Van den Berghe, G. and Luyten, F. 2014. Humanized culture of periosteal progenitors in allogeneic serum enhances osteogenic differentiation and in vivo bone formation. Stem Cells Translational Medicine. 3 (2), pp. 218-28. https://doi.org/10.5966/sctm.2012-0137
Perioperative blood transfusion is associated with a gene transcription profile characteristic of immunosuppression: a prospective cohort study
Fragkou, P., Torrance, H., Pearse, R., Ackland, G., Prowle, J., Owen, H., Hinds, C. and O'Dwyer, M. 2014. Perioperative blood transfusion is associated with a gene transcription profile characteristic of immunosuppression: a prospective cohort study. Critical Care. 18 (5). https://doi.org/10.1186/s13054-014-0541-x
The MyD88+ phenotype is an adverse prognostic factor in epithelial ovarian cancer
D’Adhemar, C., Spillane, C., Gallagher, M., Bates, M., Costello, K., Barry-O'Crowley, J., Haley, K., Kernan, N., Murphy, C., Smyth, P., O'Byrne, K., Pennington, S., Cooke, A., Ffrench, B., Martin, C., O'Donnell, D., Hennessy, B., Stordal, B., Finn, S., McCann, A., Gleeson, N., D'Arcy, T., Flood, B., O'Neill, L., Sheils, O., O'Toole, S. and O'Leary, J. 2014. The MyD88+ phenotype is an adverse prognostic factor in epithelial ovarian cancer. PLoS ONE. 9 (6), pp. 1-15. https://doi.org/10.1371/journal.pone.0100816
OvMark: a user-friendly system for the identification of prognostic biomarkers in publically available ovarian cancer gene expression datasets
Madden, S., Clarke, C., Stordal, B., Carey, M., Broaddus, R., Gallagher, W., Crown, J., Mills, G. and Hennessy, B. 2014. OvMark: a user-friendly system for the identification of prognostic biomarkers in publically available ovarian cancer gene expression datasets. Molecular Cancer. 13 (1), pp. 1-11. https://doi.org/10.1186/1476-4598-13-241
PARP inhibitors as P-glyoprotein substrates
Lawlor, D., Martin, P., Busschots, S., Thery, J., O'Leary, J.J., Hennessy, B.T. and Stordal, B. 2014. PARP inhibitors as P-glyoprotein substrates. Journal of Pharmaceutical Sciences. 103 (6), pp. 1913-1920. https://doi.org/10.1002/jps.23952
Collateral sensitivity to cisplatin in KB-8-5-11 drug-resistant cancer cells
Doherty, B., Lawlor, D., Gillet, J., Gottesman, M., O'Leary, J. and Stordal, B. 2014. Collateral sensitivity to cisplatin in KB-8-5-11 drug-resistant cancer cells. Anticancer Research. 34 (1), pp. 503-507.
In vitro development of chemotherapy and targeted therapy drug-resistant cancer cell lines: a practical guide with case studies
McDermott, M., Eustace, A., Busschots, S., Breen, L., Crown, J., Clynes, M., O'Donovan, N. and Stordal, B. 2014. In vitro development of chemotherapy and targeted therapy drug-resistant cancer cell lines: a practical guide with case studies. Frontiers in Oncology. 4, pp. 1-16. https://doi.org/10.3389/fonc.2014.00040
Effect of photodynamic therapy on proliferation and apoptosis of 3T3 fibroblasts and HeLa cells
Jankowska, A., Wiecek, P. and Burczynska, B. 2008. Effect of photodynamic therapy on proliferation and apoptosis of 3T3 fibroblasts and HeLa cells. Photomedicine and Laser Surgery. 26 (4), pp. 343-347. https://doi.org/10.1089/pho.2007.2199
Stable knockdown of hCGβ mRNA expression in bladder cancer cells results in significant inhibition of growth
Burczynska, B., Booth, M., Iles, R., Shah, A., Shiled, A. and Butler, S. 2013. Stable knockdown of hCGβ mRNA expression in bladder cancer cells results in significant inhibition of growth. Anticancer Research. 33 (9), pp. 3611-3614.
Reduction of human chorionic gonadotropin beta subunit expression by modified U1 snRNA caused apoptosis in cervical cancer cells
Jankowska, A., Gunderson, S., Andrusiewicz, M., Burczynska, B., Szczerba, A., Jarmolowski, A., Nowak-Markwitz, E. and Warchol, J. 2008. Reduction of human chorionic gonadotropin beta subunit expression by modified U1 snRNA caused apoptosis in cervical cancer cells. Molecular Cancer. 7 (26). https://doi.org/10.1186/1476-4598-7-26
Enhanced immunoreceptor tyrosine-based activation motif signaling is related to pathological bone resorption during critical illness
Vanhees, I., Gunst, J., Janssens, T., Wauters, A., Van Herck, E., Van Cromphaut, S., Van den Berghe, G. and Owen, H. 2013. Enhanced immunoreceptor tyrosine-based activation motif signaling is related to pathological bone resorption during critical illness. Hormone and Metabolic Research. 45 (12), pp. 862-9. https://doi.org/10.1055/s-0033-1351290
BRCA1/2 mutation analysis in 41 ovarian cell lines reveals only one functionally deleterious BRCA1 mutation
Stordal, B., Timms, K., Farrelly, A., Gallagher, D., Busschots, S., Renaud, M., Thery, J., Williams, D., Potter, J., Tran, T., Korpanty, G., Cremona, M., Carey, M., Li, J., Li, Y., Aslan, O., O'Leary, J., Mills, G. and Hennessy, B. 2013. BRCA1/2 mutation analysis in 41 ovarian cell lines reveals only one functionally deleterious BRCA1 mutation. Molecular Oncology. 7 (3), pp. 567-79. https://doi.org/10.1016/j.molonc.2012.12.007
Critical illness-related bone loss is associated with osteoclastic and angiogenic abnormalities
Owen, H., Vanhees, I., Solie, L., Roberts, S., Wauters, A., Luyten, F., Van Cromphaut, S. and Van den Berghe, G. 2012. Critical illness-related bone loss is associated with osteoclastic and angiogenic abnormalities. Journal of Bone and Mineral Research. 27 (7), pp. 1541-1552. https://doi.org/10.1002/jbmr.1612
Resistance to paclitaxel in a cisplatin-resistant ovarian cancer cell line is mediated by P-glycoprotein
Stordal, B., Hamon, M., McEneaney, V., Roche, S., Gillet, J., O'Leary, J., Gottesman, M. and Clynes, M. 2012. Resistance to paclitaxel in a cisplatin-resistant ovarian cancer cell line is mediated by P-glycoprotein. PLoS ONE. 7 (7), pp. 1-13. https://doi.org/10.1371/journal.pone.0040717
Hypermethylation of CpG islands and shores around specific microRNAs and mirtrons is associated with the phenotype and presence of bladder cancer
Dudziec, E., Miah, S., Choudhry, H., Owen, H., Blizard, S., Glover, M., Hamdy, F. and Catto, J. 2011. Hypermethylation of CpG islands and shores around specific microRNAs and mirtrons is associated with the phenotype and presence of bladder cancer. Clinical Cancer Research. 17 (6), pp. 1287-1296. https://doi.org/10.1158/1078-0432.CCR-10-2017
Erlotinib or gefitinib for the treatment of relapsed platinum pretreated non-small cell lung cancer and ovarian cancer: a systematic review
Murphy, M. and Stordal, B. 2011. Erlotinib or gefitinib for the treatment of relapsed platinum pretreated non-small cell lung cancer and ovarian cancer: a systematic review. Drug Resistance Updates. 14 (3), pp. 177-190. https://doi.org/10.1016/j.drup.2011.02.004
Platelet adhesion and degranulation induce pro-survival and pro-angiogenic signalling in ovarian cancer cells
Egan, K., Crowley, D., Smyth, P., O'Toole, S., Spillane, C., Martin, C., Gallagher, M., Canney, A., Norris, L., Conlon, N., McEvoy, L., Ffrench, B., Stordal, B., Keegan, H., Finn, S., McEneaney, V., Laios, A., Ducrée, J., Dunne, E., Smith, L., Berndt, M., Sheils, O., Kenny, D. and O'Leary, J. 2011. Platelet adhesion and degranulation induce pro-survival and pro-angiogenic signalling in ovarian cancer cells. PLoS ONE. 6 (10). https://doi.org/10.1371/journal.pone.0026125
Low frequency of epigenetic events in urothelial tumours from young patients
Catto, J., Owen, H., Giedl, J., Wild, P., Fine, S., Epstein, J., Humphrey, P., Dehner, L., Amin, M., Blaszyk, H., Hughes, D., Hartmann, A. and Stoehr, R. 2010. Low frequency of epigenetic events in urothelial tumours from young patients. European Urology Supplements. 9 (2), pp. 260-261. https://doi.org/10.1016/S1569-9056(10)60798-1
Low frequency of epigenetic events in urothelial tumors in young patients
Owen, H., Giedl, J., Wild, P., Fine, S., Humphrey, P., Dehner, L., Amin, M., Epstein, J., Blaszyk, H., Hughes, D., Hartmann, A., Stoehr, R. and Catto, J. 2010. Low frequency of epigenetic events in urothelial tumors in young patients. The Journal of Urology. 184 (2), pp. 459-463. https://doi.org/10.1016/j.juro.2010.03.131
Rod outer segment membrane guanylate cyclase type 1 (ROS-GC1) calcium-modulated transduction system in the sperm
Jankowska, A., Burczynska, B., Duda, T. and Warchol, J. 2010. Rod outer segment membrane guanylate cyclase type 1 (ROS-GC1) calcium-modulated transduction system in the sperm. Fertility and Sterility. 93 (3), pp. 904-912. https://doi.org/10.1016/j.fertnstert.2008.10.048
Chondrocyte p21(WAF1/CIP1) expression is increased by dexamethasone but does not contribute to dexamethasone-induced growth retardation in vivo
Owen, H., Ahmed, S. and Farquharson, C. 2009. Chondrocyte p21(WAF1/CIP1) expression is increased by dexamethasone but does not contribute to dexamethasone-induced growth retardation in vivo. Calcified Tissue International and Musculoskeletal Research. 85 (4), pp. 326-334. https://doi.org/10.1007/s00223-009-9276-0
Distinct microRNA alterations characterize high- and low-grade bladder cancer
Catto, J., Miah, S., Owen, H., Bryant, H., Myers, K., Dudziec, E., Larré, S., Milo, M., Rehman, I., Rosario, D., Di Martino, E., Knowles, M., Meuth, M., Harris, A. and Hamdy, F. 2009. Distinct microRNA alterations characterize high- and low-grade bladder cancer. Cancer Research. 69 (21), pp. 8472-8481. https://doi.org/10.1158/0008-5472.CAN-09-0744
ERCC1 expression and RAD51B activity correlate with cell cycle response to platinum drug treatment not DNA repair
Stordal, B. and Davey, R. 2009. ERCC1 expression and RAD51B activity correlate with cell cycle response to platinum drug treatment not DNA repair. Cancer Chemotherapy and Pharmacology. 63 (4), pp. 661-672. https://doi.org/10.1007/s00280-008-0783-x
Citations, citations everywhere but did anyone read the paper? [Letter to the Editor]
Stordal, B. 2009. Citations, citations everywhere but did anyone read the paper? [Letter to the Editor]. Colloids and surfaces. B, Biointerfaces. 72 (2), pp. 312 -312. https://doi.org/10.1016/j.colsurfb.2009.04.001
A systematic review of genes involved in the inverse resistance relationship between cisplatin and paclitaxel chemotherapy: role of BRCA1
Stordal, B. and Davey, R. 2009. A systematic review of genes involved in the inverse resistance relationship between cisplatin and paclitaxel chemotherapy: role of BRCA1. Current Cancer Drug Targets. 9 (3), pp. 354-365. https://doi.org/10.2174/156800909788166592
Dexamethasone-induced expression of the glucocorticoid response gene lipocalin 2 in chondrocytes
Owen, H., Roberts, S., Ahmed, S. and Farquharson, C. 2008. Dexamethasone-induced expression of the glucocorticoid response gene lipocalin 2 in chondrocytes. American Journal of Physiology - Endocrinology and Metabolism. 294 (6), pp. E1023-E1034. https://doi.org/10.1152/ajpendo.00586.2007
Identification of a novel splice variant of the haloacid dehalogenase: PHOSPHO1
Roberts, S., Owen, H. and Farquharson, C. 2008. Identification of a novel splice variant of the haloacid dehalogenase: PHOSPHO1. Biochemical and Biophysical Research Communications. 371 (4), pp. 872-876. https://doi.org/10.1016/j.bbrc.2008.04.163
Platinum resistance needs the mythbusters [Letter to the Editor]
Stordal, B. and Davey, R. 2008. Platinum resistance needs the mythbusters [Letter to the Editor]. Toxicology letters. 180 (3), p. 230. https://doi.org/10.1016/j.toxlet.2008.07.003
A 39 kDa fragment of endogenous ASK1 suggests specific cleavage not degradation by the proteasome
Stordal, B. and Davey, R. 2008. A 39 kDa fragment of endogenous ASK1 suggests specific cleavage not degradation by the proteasome. IUBMB Life. 60 (3), pp. 180-184. https://doi.org/10.1002/iub.24
Identification and functional involvement of lipocalin 2 in glucocorticoid-induced growth retardation
Owen, H., Ahmed, S. and Farquharson, C. 2007. Identification and functional involvement of lipocalin 2 in glucocorticoid-induced growth retardation. 29th Annual Meeting of the American Society for Bone and Mineral Research. Honolulu, HI 16 - 19 Sep 2007 Wiley.
The role of P21CIP1/WAF1 in glucocorticoid induced growth retardation
Owen, H., Ahmed, S. and Farquharson, C. 2007. The role of P21CIP1/WAF1 in glucocorticoid induced growth retardation. 2007 Annual Meeting of the Bone Research Society. Southampton, UK 03 - 05 Jul 2007 Wiley.
Identification of lipocalin 2, a novel glucocorticoid responsive gene in growth plate chondrocytes
Owen, H., Ahmed, S. and Farquharson, C. 2007. Identification of lipocalin 2, a novel glucocorticoid responsive gene in growth plate chondrocytes. 2007 Annual Meeting of the Bone Research Society. Southampton, UK 03 - 05 Jul 2007 Wiley.
The growth plate sparing effects of the selective glucocorticoid receptor modulator, AL-438
Owen, H., Miner, J., Ahmed, S. and Farquharson, C. 2007. The growth plate sparing effects of the selective glucocorticoid receptor modulator, AL-438. Molecular and Cellular Endocrinology. 264 (1-2), pp. 164-70. https://doi.org/10.1016/j.mce.2006.11.006
A systematic review of platinum and taxane resistance from bench to clinic: an inverse relationship
Stordal, B., Pavlakis, N. and Davey, R. 2007. A systematic review of platinum and taxane resistance from bench to clinic: an inverse relationship. Cancer Treatment Reviews. 33 (8), pp. 688-703. https://doi.org/10.1016/j.ctrv.2007.07.013
Understanding cisplatin resistance using cellular models
Stordal, B. and Davey, M. 2007. Understanding cisplatin resistance using cellular models. IUBMB Life. 59 (11), pp. 696-699. https://doi.org/10.1080/15216540701636287
Oxaliplatin for the treatment of cisplatin-resistant cancer: a systematic review
Stordal, B., Pavlakis, N. and Davey, R. 2007. Oxaliplatin for the treatment of cisplatin-resistant cancer: a systematic review. Cancer Treatment Reviews. 33 (4), pp. 347-357. https://doi.org/10.1016/j.ctrv.2007.01.009
Glucocorticoids stimulate p21 expression in growth plate chondrocytes
Owen, H., Ahmed, S. and Farquharson, C. 2006. Glucocorticoids stimulate p21 expression in growth plate chondrocytes. 2006 Annual Meeting of the Bone Research Society. Southampton, UK 05 - 06 Jul 2006 Wiley.
Glucocorticoids induce lipocalin 2 gene expression in growth plate chondrocytes.
Owen, H., Ahmed, S. and Farquharson, C. 2006. Glucocorticoids induce lipocalin 2 gene expression in growth plate chondrocytes. 28th Annual Meeting of the American Society for Bone and Mineral Research. Philadelphia, PA 15 - 19 Sep 2006 Wiley.
The role of the 11 beta HSD shuttle in modulating the effects of proinflammatory cytokines on the growth plate
MacRae, V., Owen, H., Ahmed, S. and Farquharson, C. 2006. The role of the 11 beta HSD shuttle in modulating the effects of proinflammatory cytokines on the growth plate. 28th Annual Meeting of the American Society for Bone and Mineral Research. Philadelphia, PA 15 - 19 Sep 2006 Wiley.
Similar chromosomal changes in cisplatin and oxaliplatin-resistant sublines of the H69 SCLC cell line are not associated with platinum resistance
Stordal, B., Peters, G. and Davey, R. 2006. Similar chromosomal changes in cisplatin and oxaliplatin-resistant sublines of the H69 SCLC cell line are not associated with platinum resistance. Genes, Chromosomes & Cancer. 45 (12), pp. 1094-105. https://doi.org/10.1002/gcc.20373
Oxaliplatin induces drug resistance more rapidly than cisplatin in H69 small cell lung cancer cells
Stordal, B., Davey, M. and Davey, R. 2006. Oxaliplatin induces drug resistance more rapidly than cisplatin in H69 small cell lung cancer cells. Cancer Chemotherapy and Pharmacology. 58 (2), pp. 256-265. https://doi.org/10.1007/s00280-005-0148-7