Mechanisms of cisplatin-induced cell death in malignant mesothelioma cells: Role of inhibitor of apoptosis proteins (IAPs) and caspases

Cregan,Inez  L.; Dharmarajan,Arun  M.; Fox,Simon  A.

doi:10.3892/ijo.2012.1715

Mechanisms of cisplatin-induced cell death in malignant mesothelioma cells: Role of inhibitor of apoptosis proteins (IAPs) and caspases

Authors:
- Inez L. Cregan
- Arun M. Dharmarajan
- Simon A. Fox
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Affiliations: School of Anatomy and Human Biology, The University of Western Australia, Crawley, WA, Australia, Molecular Pharmacology Laboratory, School of Pharmacy, Western Australian Biomedical Research Institute & Curtin Health Innovation Research Institute, Curtin University, Bentley, WA, Australia
Published online on: November 28, 2012 https://doi.org/10.3892/ijo.2012.1715
Pages: 444-452
Copyright: © Cregan et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Malignant mesothelioma (MM) is an aggressive and highly chemoresistant tumour. Although cisplatin is used in frontline therapy of this disease treatment remains palliative at best. The biochemical pathways activated by cisplatin and the mechanisms of resistance in mesothelioma cells are poorly understood. Overexpression of inhibitor of apoptosis proteins (IAPs) has been described in clinical mesothelioma tumours and proposed as therapeutic targets. In this study, we examined cisplatin-induced cell death pathways and IAPs in three mesothelioma-derived cell lines. Cisplatin induced cell death in mesothelioma cell lines was characterised by biochemical mechanisms classically associated with apoptosis including: mitochondrial depolarisation, phosphatidylserine translocation and caspase activation. Surprisingly mRNA expression of IAPs in mesothelioma was not upregulated relative to primary mesothelial cells except for survivin which was higher in the most resistant cell line. In contrast, protein expression of both XIAP and survivin was upregulated in all mesothelioma cells, consistent with post-translational regulation. Knockdown of either XIAP or survivin by RNAi did not affect the sensitivity to cisplatin in any of the cell lines. Survivin RNAi did, however, inhibit proliferation in the highest expressing cell line, ONE58. The pan-caspase inhibitor z-VAD and the more selective caspase 3/7 inhibitor z-DEVD had no effect upon the sensitivity of any of the cell lines to cisplatin indicating that caspase-independent pathways predominate. The findings of the present study provide insights into cisplatin-induced mechanisms in mesothelioma cells and show that alternative pathways are operating which may provide new options for targeting this extremely resistant tumour.

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1.	Spugnini EP, Bosari S, Citro G, Lorenzon I, Cognetti F and Baldi A: Human malignant mesothelioma: molecular mechanisms of pathogenesis and progression. Int J Biochem Cell Biol. 38:2000–2004. 2006. View Article : Google Scholar : PubMed/NCBI
2.	Ray M and Kindler HL: Malignant pleural mesothelioma: an update on biomarkers and treatment. Chest. 136:888–896. 2009. View Article : Google Scholar : PubMed/NCBI
3.	Rabik CA and Dolan ME: Molecular mechanisms of resistance and toxicity associated with platinating agents. Cancer Treat Rev. 33:9–23. 2007. View Article : Google Scholar : PubMed/NCBI
4.	Koberle B, Tomicic MT, Usanova S and Kaina B: Cisplatin resistance: preclinical findings and clinical implications. Biochim Biophys Acta. 1806:172–182. 2010.PubMed/NCBI
5.	Davidson B: Expression of cancer-associated molecules in malignant mesothelioma. Biomark Insights. 2:173–184. 2007.PubMed/NCBI
6.	Kleinberg L, Lie AK, Florenes VA, Nesland JM and Davidson B: Expression of inhibitor-of-apoptosis protein family members in malignant mesothelioma. Hum Pathol. 38:986–994. 2007. View Article : Google Scholar : PubMed/NCBI
7.	Zaffaroni N, Costa A, Pennati M, et al: Survivin is highly expressed and promotes cell survival in malignant peritoneal mesothelioma. Cell Oncol. 29:453–466. 2007.PubMed/NCBI
8.	Fox SA, Kusmiaty, Loh SS, Dharmarajan AM and Garlepp MJ: Cisplatin and TNF-alpha downregulate transcription of Bcl-xL in murine malignant mesothelioma cells. Biochem Biophys Res Commun. 337:983–991. 2005. View Article : Google Scholar : PubMed/NCBI
9.	Manning LS, Whitaker D, Murch AR, et al: Establishment and characterization of five human malignant mesothelioma cell lines derived from pleural effusions. Int J Cancer. 47:285–290. 1991. View Article : Google Scholar : PubMed/NCBI
10.	Altieri DC: Survivin and IAP proteins in cell-death mechanisms. Biochem J. 430:199–205. 2010. View Article : Google Scholar : PubMed/NCBI
11.	Rozen S and Skaletsky H: Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol. 132:365–386. 2000.PubMed/NCBI
12.	Vandesompele J, De Preter K, Pattyn F, et al: Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 3:Research 0034. 2002. View Article : Google Scholar : PubMed/NCBI
13.	Matsumiya T, Imaizumi T, Yoshida H, Kimura H and Satoh K: Cisplatin inhibits the expression of X-chromosome-linked inhibitor of apoptosis protein in an oral carcinoma cell line. Oral Oncol. 37:296–300. 2001. View Article : Google Scholar : PubMed/NCBI
14.	Nomura T, Mimata H, Yamasaki M and Nomura Y: Cisplatin inhibits the expression of X-linked inhibitor of apoptosis protein in human LNCaP cells. Urol Oncol. 22:453–460. 2004. View Article : Google Scholar : PubMed/NCBI
15.	Siddik ZH: Cisplatin: mode of cytotoxic action and molecular basis of resistance. Oncogene. 22:7265–7279. 2003. View Article : Google Scholar : PubMed/NCBI
16.	Andjilani M, Droz JP, Benahmed M and Tabone E: Down-regulation of FAK and IAPs by laminin during cisplatin-induced apoptosis in testicular germ cell tumors. Int J Oncol. 28:535–542. 2006.PubMed/NCBI
17.	Golovine K, Makhov P, Uzzo RG, et al: Cadmium down-regulates expression of XIAP at the post-transcriptional level in prostate cancer cells through an NF-kappaB-independent, proteasome-mediated mechanism. Mol Cancer. 9:1832010. View Article : Google Scholar
18.	Zhao J, Tenev T, Martins LM, Downward J and Lemoine NR: The ubiquitin-proteasome pathway regulates survivin degradation in a cell cycle-dependent manner. J Cell Sci. 113(Pt 23): 4363–4371. 2000.PubMed/NCBI
19.	Altieri DC: New wirings in the survivin networks. Oncogene. 27:6276–6284. 2008. View Article : Google Scholar : PubMed/NCBI
20.	Holcik M: Translational upregulation of the X-linked inhibitor of apoptosis. Ann N Y Acad Sci. 1010:249–258. 2003. View Article : Google Scholar : PubMed/NCBI
21.	Li Y, Jian Z, Xia K, et al: XIAP is related to the chemoresistance and inhibited its expression by RNA interference sensitize pancreatic carcinoma cells to chemotherapeutics. Pancreas. 32:288–296. 2006. View Article : Google Scholar : PubMed/NCBI
22.	Zhang Y, Wang Y, Gao W, et al: Transfer of siRNA against XIAP induces apoptosis and reduces tumor cells growth potential in human breast cancer in vitro and in vivo. Breast Cancer Res Treat. 96:267–277. 2006. View Article : Google Scholar : PubMed/NCBI
23.	Mita AC, Mita MM, Nawrocki ST and Giles FJ: Survivin: key regulator of mitosis and apoptosis and novel target for cancer therapeutics. Clin Cancer Res. 14:5000–5005. 2008. View Article : Google Scholar : PubMed/NCBI
24.	Liu WS, Yan HJ, Qin RY, et al: siRNA directed against survivin enhances pancreatic cancer cell gemcitabine chemosensitivity. Dig Dis Sci. 54:89–96. 2009. View Article : Google Scholar : PubMed/NCBI
25.	Yamaguchi Y, Shiraki K, Fuke H, et al: Targeting of X-linked inhibitor of apoptosis protein or survivin by short interfering RNAs sensitize hepatoma cells to TNF-related apoptosis-inducing ligand- and chemotherapeutic agent-induced cell death. Oncol Rep. 14:1311–1316. 2005.
26.	Hopkins-Donaldson S, Belyanskaya LL, Simoes-Wust AP, et al: p53-induced apoptosis occurs in the absence of p14(ARF) in malignant pleural mesothelioma. Neoplasia. 8:551–559. 2006. View Article : Google Scholar : PubMed/NCBI
27.	Cummings BS, Kinsey GR, Bolchoz LJ and Schnellmann RG: Identification of caspase-independent apoptosis in epithelial and cancer cells. J Pharmacol Exp Ther. 310:126–134. 2004. View Article : Google Scholar : PubMed/NCBI
28.	Kim R, Emi M and Tanabe K: Caspase-dependent and -independent cell death pathways after DNA damage (Review). Oncol Rep. 14:595–599. 2005.PubMed/NCBI
29.	Liu L, Xing D and Chen WR: Micro-calpain regulates caspase-dependent and apoptosis inducing factor-mediated caspase-independent apoptotic pathways in cisplatin-induced apoptosis. Int J Cancer. 125:2757–2766. 2009. View Article : Google Scholar
30.	Lock EA, Reed CJ, Kinsey GR and Schnellmann RG: Caspase-dependent and -independent induction of phosphatidylserine externalization during apoptosis in human renal carcinoma Cak(1)-1 and A-498 cells. Toxicology. 229:79–90. 2007. View Article : Google Scholar
31.	Delavallee L, Cabon L, Galan-Malo P, Lorenzo HK and Susin SA: AIF-mediated caspase-independent necroptosis: a new chance for targeted therapeutics. IUBMB Life. 63:221–232. 2011. View Article : Google Scholar : PubMed/NCBI