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Cancer Chemotherapy and Pharmacology
Published in: Cancer Chemotherapy and Pharmacology 2/2012

01-02-2012 | Original Article

Inhibition of glutathione synthesis reverses Krüppel-like factor 4-mediated cisplatin resistance

Authors: Yongsheng Jia, Wenjian Zhang, Honglin Liu, Liang Peng, Zhihua Yang, Jinning Lou

Published in: Cancer Chemotherapy and Pharmacology | Issue 2/2012

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Abstract

Purpose

To explore the protective effect of KLF4 against cytotoxicity induced by cisplatin and its possible mechanisms.

Methods

The expression levels of KLF4 were detected by RT-PCR and western blot in cancer stem-like cells derived from hepatocarcinoma (T3A-A3) and the hepatocarcinoma cell line HepG2. KLF4 was knocked down in T3A-A3 by infection of pLVTHM-shKLF4 lentivirus and ectopic expressed in HepG2 by infection of pWPTS-KLF4 lentivirus. The MTT assay was carried out to determine the impact of KLF4 on cell survival in response to cisplatin. Cisplatin-induced DNA damage was measured by TUNEL staining. Glutathione content was measured by enzymatic assay. Buthionine sulfoximine was used to deplete the content of glutathione. The expression of γ-glutamylcysteine synthetase was analyzed by RT-PCR in HepG2 cells ectopic expressed KLF4.

Results

With a higher level of KLF4, T3A-A3 cells were found to be more resistant to cisplatin than HepG2 cells. KLF4 knockdown was found to reduce cisplatin resistance in T3A-A3 cells. Ectopic expression of KLF4 in HepG2 cells was found to be associated with heightened resistance to DNA damage after exposure to cisplatin. Furthermore, the content of glutathione was found to be higher in T3A-A3 cells than in HepG2 cells. A nearly twofold increase in the cellular level of glutathione was identified in HepG2 cells with ectopic expression of KLF4. This was accompanied by heightened resistance to cisplatin. KLF4-mediated resistance to cisplatin in HepG2 cells was found to be completely abrogated by treatment with buthionine sulfoximine, an inhibitor of glutathione synthesis, which did not affect the expression of KLF4. Moreover, the mRNA expression of γ-glutamylcysteine synthetase, a rate-limiting enzyme of glutathione synthesis was up-regulated by KLF.

Conclusion

We conclude that KLF4 regulates the cellular sensitivity to cisplatin in hepatocarcinoma stem-like cells and hepatocarcinoma cells by elevating intracellular glutathione levels.
Literature
1.
An Y, Ongkeko WM (2009) ABCG2: the key to chemoresistance in cancer stem cells? Expert Opin Drug Metab Toxicol 5:1529–1542PubMedCrossRef
2.
Fais S (2011) Proton pump inhibitor-induced tumour cell death by inhibition of a detoxification mechanism. J Intern Med 267:515–525CrossRef
3.
Huang G, Mills L, Worth LL (2007) Expression of human glutathione S-transferase P1 mediates the chemosensitivity of osteosarcoma cells. Mol Cancer Ther 6:1610–1619PubMedCrossRef
4.
Varin E, Denoyelle C, Brotin E, Meryet-Figuiere M, Giffard F, Abeilard E, Goux D, Gauduchon P, Icard P, Poulain L (2010) Downregulation of Bcl-xL and Mcl-1 is sufficient to induce cell death in mesothelioma cells highly refractory to conventional chemotherapy. Carcinogenesis 31:984–993PubMedCrossRef
5.
Ott K, Rachakonda PS, Panzram B, Keller G, Lordick F, Becker K, Langer R, Buechler M, Hemminki K, Kumar R (2011) DNA repair gene and MTHFR gene polymorphisms as prognostic markers in locally advanced adenocarcinoma of the esophagus or stomach treated with cisplatin and 5-fluorouracil-based neoadjuvant chemotherapy. Ann Surg Oncol [Epub ahead of print]
6.
Wang GuoZ (2007) The role of sulfur in platinum anticancer chemotherapy. Anticancer Agents Med Chem 7:19–34CrossRef
7.
Godwin AK, Meister A, O’Dwyer PJ, Huang CS, Hamilton TC, Anderson ME (1992) High resistance to cisplatin in human ovarian cancer cell lines is associated with marked increase of glutathione synthesis. Proc Natl Acad Sci USA 89:3070–3074PubMedCrossRef
8.
Zhang K, Chew M, Yang EB, Wong KP, Mack P (2001) Modulation of cisplatin cytotoxicity and cisplatin-induced DNA cross-links in HepG2 cells by regulation of glutathione-related mechanisms. Mol Pharmacol 59:837–843PubMed
9.
Anderson ME, Naganuma A, Meister A (1990) Protection against cisplatin toxicity by administration of glutathione ester. FASEB J 4:3251–3255PubMed
10.
Yang Z, Faustino PJ, Andrews PA, Monastra R, Rasmussen AA, Ellison CD, Cullen KJ (2000) Decreased cisplatin/DNA adduct formation is associated with cisplatin resistance in human head and neck cancer cell lines. Cancer Chemother Pharmacol 46:255–262PubMedCrossRef
11.
Alison MR, Lim SM, Nicholson LJ (2010) Cancer stem cells: problems for therapy? J Pathol 223:147–161PubMed
12.
13.
Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126:663–676PubMedCrossRef
14.
Yu F, Li J, Chen H, Fu J, Ray S, Huang S, Zheng H, Ai W (2011) Kruppel-like factor 4 (KLF4) is required for maintenance of breast cancer stem cells and for cell migration and invasion. Oncogene 30:1–12
15.
Rowland BD, Bernards R, Peeper DS (2005) The KLF4 tumour suppressor is a transcriptional repressor of p53 that acts as a context-dependent oncogene. Nat Cell Biol 7:1074–1082PubMedCrossRef
16.
Wu LQ, Zhang WJ, Niu JX, Ye LY, Yang ZH, Grau GE, Lou JN (2008) Phenotypic and functional differences between human liver cancer endothelial cells and liver sinusoidal endothelial cells. J Vasc Res 45:78–86PubMedCrossRef
17.
Xu N, Papagiannakopoulos T, Pan G, Thomson JA, Kosik KS (2009) MicroRNA-145 regulates OCT4, SOX2, and KLF4 and represses pluripotency in human embryonic stem cells. Cell 137:647–658PubMedCrossRef
18.
Tietze F (1969) Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. Anal Biochem 27:502–522PubMedCrossRef
19.
Chen HH, Kuo MT (2010) Role of glutathione in the regulation of Cisplatin resistance in cancer chemotherapy. Met Based Drugs 2010.pii:430939
20.
Forman HJ, Zhang H, Rinna A (2009) Glutathione: overview of its protective roles, measurement, and biosynthesis. Mol Aspects Med 30:1–12PubMedCrossRef
21.
Dudley AC, Khan ZA, Shih SC, Kang SY, Zwaans BM, Bischoff J, Klagsbrun M (2008) Calcification of multipotent prostate tumor endothelium. Cancer Cell 14:201–211PubMedCrossRef
22.
Reya T, Morrison SJ, Clarke MF, Weissman IL (2001) Stem cells, cancer, and cancer stem cells. Nature 414:105–111PubMedCrossRef
23.
Wong CW, Hou PS, Tseng SF, Chien CL, Wu KJ, Chen HF, Ho HN, Kyo S, Teng SC (2010) Kruppel-like transcription factor 4 contributes to maintenance of telomerase activity in stem cells. Stem Cells 28:1510–1517PubMedCrossRef
24.
Zhou Q, Hong Y, Zhan Q, Shen Y, Liu Z (2009) Role for Kruppel-like factor 4 in determining the outcome of p53 response to DNA damage. Cancer Res 69:8284–8292PubMedCrossRef
25.
Foster KW, Liu Z, Nail CD, Li X, Fitzgerald TJ, Bailey SK, Frost AR, Louro ID, Townes TM, Paterson AJ, Kudlow JE, Lobo-Ruppert SM, Ruppert JM (2005) Induction of KLF4 in basal keratinocytes blocks the proliferation-differentiation switch and initiates squamous epithelial dysplasia. Oncogene 24:1491–1500PubMedCrossRef
26.
Rodriguez-Garcia ME, Quiroga AG, Castro J, Ortiz A, Aller P, Mata F (2009) Inhibition of p38-MAPK potentiates cisplatin-induced apoptosis via GSH depletion and increases intracellular drug accumulation in growth-arrested kidney tubular epithelial cells. Toxicol Sci 111:413–423PubMedCrossRef
27.
Ishimoto T, Nagano O, Yae T, Tamada M, Motohara T, Oshima H, Oshima M, Ikeda T, Asaba R, Yagi H, Masuko T, Shimizu T, Ishikawa T, Kai K, Takahashi E, Imamura Y, Baba Y, Ohmura M, Suematsu M, Baba H, Saya H (2011) CD44 Variant Regulates Redox Status in Cancer Cells by Stabilizing the xCT Subunit of System xc(−) and Thereby Promotes Tumor Growth. Cancer Cell 19:387–400
28.
Noctor G, Gomez L, Vanacker H, Foyer CH (2002) Interactions between biosynthesis, compartmentation and transport in the control of glutathione homeostasis and signalling. J Exp Bot 53:1283–1304PubMedCrossRef
Metadata
Title
Inhibition of glutathione synthesis reverses Krüppel-like factor 4-mediated cisplatin resistance
Authors
Yongsheng Jia
Wenjian Zhang
Honglin Liu
Liang Peng
Zhihua Yang
Jinning Lou
Publication date
01-02-2012
Publisher
Springer-Verlag
Published in
Cancer Chemotherapy and Pharmacology / Issue 2/2012
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI
https://doi.org/10.1007/s00280-011-1708-7

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