Mechanism of cross‐resistance to cisplatin in a mitomycin C‐resistant human bladder cancer cell line

Singh, Shivendra V.; Xu, Baocheng; Jani, Jitesh P.; Emerson, Erling O.; Backes, Mary G.; Rihn, Christopher; Scalamogna, Domenic; Stemmler, Nancy; Specht, Susan; Blanock, Kurt; Katoh, Arthur; Gupta, V. D.

doi:10.1002/ijc.2910610326

Cited by 19 publications

(9 citation statements)

References 17 publications

(30 reference statements)

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“…While GST activity was higher by about 1.5-fold in the J82/MMC-2 cell line compared with J82/WT ( p < 0.05), this activity did not differ significantly between J82/MMC and J82/MMC-2 cells. We have shown previously that .rr-type GST protein is the major isoenzyme in J82/WT and J82/MMC cells (Singh et al, 1995). Immunoblot analysis for GST .rr expression revealed that the elevated GST activity in the J82/MMC-2 cell line was due to an increase in the level of GST .rr-type protein (Fig.…”

Section: Resultsmentioning

confidence: 68%

“…Total RNA from J82/WT, J82/MMC or J82/MMC-2 cells (lo7) was isolated by acid-guanidinium-thiocyanate-phenol-chloroform extraction as described previously (Singh et al, 1995). The integrity of the RNA was assessed by ethidium bromide staining of fractionated RNA in agarose gels.…”

Section: Northern Blot Analysismentioning

confidence: 99%

“…GST activity was measured using the method described by Habig et al (1974). Immunoblot analysis for expression of GST IT, using equal amounts of 14,000 g supernatant fraction protein from J82/WT, J82/MMC and J82/MMC-2 cells, was performed as described previously (Singh et al, 1995). Bands were visualized using a chemiluminescence kit according to the manufacturer's instructions (Alpha Diagnostics, San Antonio, TX).…”

Section: Quantitation Of Gsh and Gst Levels And Immunoblot Analysis Fmentioning

confidence: 99%

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Biochemical characterization of a mitomycin C-resistant human bladder cancer cell line

et al. 1996

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This study describes characteristics of a mitomycin C (MMC)-resistant human bladder cancer cell line, )82/MMC-2, which was established by repeated in vitro exposures of a 6-fold MMC-resistant variant (J82lMMC) to 18 nM MMC. A 9.6-fold higher concentration of MMC was required to kill 50% of the )82/MMC-2 sub-line compared with parental cells ()82/WT). NADPH cytochrome P450 reductase and DT-diaphorase activities were significantly lower in J82/MMC-2 cells compared with J82/WT, suggesting that reduced sensitivity of )82/MMC-2 cells to MMC resulted from impaired drug activation. Consistent with this hypothesis, the formation of MMC-alkylating metabolites was significantly lower in )82/MMC-2 cells compared with J82iWT. Furthermore, DT-diaphorase activity in J82/MMC-2 cells was significantly lower compared with the 6-fold MMCresistant variant. Glutathione (GSH) levels were comparable in all 3 cell lines. Although GSH transferase (GST) activity was significantly higher in the )82/MMC-2 cells compared with )82/WT, this enzyme activity did not differ between 6-and 9.6-fold MMC-resistant variants. Whereas DNA polymerase Q mRNA expression was comparable in these cell lines, levels of DNA ligase I mRNA were slightly lower in both MMC-resistant variants relative to )82/WT. However, the DNA polymerase p mRNA level was markedly higher in the J82/MMC-2 cell line compared with either )82/WT or )82/MMC. Thus, emergence of a higher level of resistance to MMC in J82/MMC-2 cells compared with J82/MMC may be attributed to (i) impaired drug activation through further reduction in DT-diaphorase activity and (ii) enhanced DNA repair through over-expression of DNA polymerase p.o 1996 Wiley-Liss, Inc.Mitomycin C (MMC), a bioreductive alkylating agent, has demonstrated activity against various malignancies, including bladder cancer (Crooke and Bradner, 1976). However, the clinical usefulness of MMC may be limited at least in part due to drug resistance (Moertel et al., 1968; Wilson et al., 1987). Several different mechanisms have been proposed to account for tumor cell resistance to MMC, including impaired drug activation due to down-regulation of an MMC bioactivation enzyme(s) (Hoban et al., 1990; Pan et al., 1992;Xu et al., 1994a), reduced drug accumulation (Dorr et al., 1987;Shibata et al., 1995), decreased MMC-induced oxygen radical formation (Dusre et al., 1990), enhanced glutathione (GSH)/GSH transferase (GST)-mediated drug inactivation Singh et al., 1992; Xu et al., 19946) and increased DNA repair (Dulhanty et al., 1989).We previously reported characterization of a human bladder cancer cell line (J82/MMC) exhibiting about 6.0-fold more resistance to MMC compared with parental cells ( Xu et al., 1994a). The J82iMMC sub-line was established by repeated continuous exposures of J82/WT cells to increasing concentrations of MMC (9-18 nM) over a period of about 10 months. Previous studies from our laboratory have suggested that cellular resistance to MMC in the J82/MMC cell line may be manifested by multiple mechanisms, including impaired...

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Section: Resultsmentioning

confidence: 68%

Section: Northern Blot Analysismentioning

confidence: 99%

Section: Quantitation Of Gsh and Gst Levels And Immunoblot Analysis Fmentioning

confidence: 99%

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Biochemical characterization of a mitomycin C-resistant human bladder cancer cell line

et al. 1996

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“…On the other hand, the overexpression of π‐type glutathione transferase, implying enhanced intracellular drug conjugation/inactivation, has been shown in a bladder cancer clone (derived from J82) which has cross‐resistance between mitomycin C and cisplatin. The latter is not a drug to which resistance is mediated by classical (P‐gp‐associated) MDR [18]. Ohga et al.…”

Section: Discussionmentioning

confidence: 99%

Cellular resistance to mitomycin C is associated with overexpression of MDR‐1 in a urothelial cancer cell line (MGH‐U1)

Hayes¹,

Birch²,

Cooper³

et al. 2001

BJU International

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Objective To compare multidrug resistance (MDR)-1 and MDR-3 gene expression in a new urothelial cancer cell line (MGHU-1, with resistance to mitomycin C) against controls and the established (epirubicinresistant) MDR clone, and to correlate MDR with cytotoxicity data. Materials and methods Resistance to mitomycin C was induced by the long-term exposure of wild-type MGHU-1 cells to increasing concentrations (20±400 nmol/L) of mitomycin C. The cytotoxicity of mitomycin C or epirubicin was then compared in MGHU-1, MGHU-MMC (mitomycin C-resistant) and MGHU-1R (established MDR) cells, using the tetrazolium biomass assay. The expression of MDR-1 and -3 was investigated by the reverse transcriptasepolymerase chain reaction, using cDNA-speci®c primers after titration, and compared with DNA and negative controls. Results MDR-1 and -3 were signi®cantly and equally overexpressed in MGHU-1R, and associated with a dramatic increase in the 50% inhibitory drug concentration (P<0.001) for mitomycin C and epirubicin against controls. In MGHU-MMC, the overexpression of MDR-1 was three times greater than that of MDR-3. The cytotoxicity pro®le for both agents was very similar to that of MGHU-1R. Trace amounts of MDR-1, but not MDR-3, were identi®ed in the MGHU-1 wild-type. Conclusions Urothelial cancer cell resistance to mitomycin C is associated with cross-resistance to epirubicin and overexpression of MDR-1, suggesting that mitomycin C falls within the MDR category. Clinical application of this methodology may allow patients to be identi®ed who are unlikely to bene®t from intravesical chemotherapy.

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“…For numerous cancer cell lines (derived from prostate, lung, ovary and cervical cancer), a correlation between MT expression and cisplatin resistance has been observed (Kasahara et al, 1991, Kondo et al, 1995, Mellish et al, 1993, Surowiak et al, 2007. For bladder cancer cell lines cisplatin resistance, was also correlated with increased levels of MT (Siegsmund et al, 1999, Singh et al, 1995. A role of MT for cisplatin resistance in bladder cancer has been proposed by Satoh and co-workers (Satoh et al, 1994).…”

Section: Introductionmentioning

confidence: 94%

The Molecular Basis of Cisplatin Resistance in Bladder Cancer Cells

Köberle¹,

Piée-Staffa²

2012

Bladder Cancer - From Basic Science to Robotic Surgery

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Mechanism of cross‐resistance to cisplatin in a mitomycin C‐resistant human bladder cancer cell line

Cited by 19 publications

References 17 publications

Biochemical characterization of a mitomycin C-resistant human bladder cancer cell line

Biochemical characterization of a mitomycin C-resistant human bladder cancer cell line

Cellular resistance to mitomycin C is associated with overexpression of MDR‐1 in a urothelial cancer cell line (MGH‐U1)

The Molecular Basis of Cisplatin Resistance in Bladder Cancer Cells

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