An etoposide-resistant lung cancer subline overexpresses the multidrug resistance-associated protein

L. A. Doyle, D. D. Ross, J. V. Ordonez, W. Yang, Y. Gao, Y. Tong, C. P. Belani, J. C. Gutheil

Research output: Contribution to journalArticle

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Abstract

We have characterised an etoposide-resistant subline of the small-cell lung cancer cell line, UMCC-1, derived at our centre. Subline UMCC-1/VP was developed by culturing the parent line in increasing concentrations of etoposide over 16 months. UMCC-1/VP is 20-fold resistant to etoposide by MTT assays, relative to the parent line, and is cross-resistant to doxorubicin, vincristine and actinomycin D, but not to taxol, cisplatin, melphalan, thiotepa or idarubicin. Topoisomerase II immunoblotting demonstrates a 50% reduction of the protein in the resistant subline. The UMCC-1/VP subline demonstrates a marked decrease in the accumulation of [3H]etoposide relative to the parent line, as well as a modest reduction in the accumulation of daunorubicin. Reverse transcription-polymerase chain reaction assays demonstrate no detectable mdr1 expression but marked expression of the multidrug resistance-associated protein (MRP) gene in the resistant subline. Northern blotting with an MRP cDNA probe confirms marked overexpression of the MRP gene only in the UMCC-1/VP subline. Western blotting with antisera against MRP peptide confirms a 195 kDa protein band in the UMCC-1/VP subline. Southern blotting experiments demonstrate a 10-fold amplification of the MRP gene in the resistant subline. Depletion of glutathione with buthionine sulphoximine sensitised UMCC-1/VP cells to daunorubicin and etoposide. Our studies indicate that MRP gene expression may be induced by etoposide and may lead to reduced accumulation of the drug.

Original languageEnglish (US)
Pages (from-to)535-542
Number of pages8
JournalBritish Journal of Cancer
Volume72
Issue number3
DOIs
StatePublished - Sep 1995

Fingerprint

Viral Structural Proteins
Multidrug Resistance-Associated Proteins
Etoposide
Lung Neoplasms
Daunorubicin
Idarubicin
Thiotepa
Genes
Buthionine Sulfoximine
Type II DNA Topoisomerase
Melphalan
Small Cell Lung Carcinoma
Dactinomycin
Vincristine
Southern Blotting
Paclitaxel
Immunoblotting
Northern Blotting
Doxorubicin
Cisplatin

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Doyle, L. A., Ross, D. D., Ordonez, J. V., Yang, W., Gao, Y., Tong, Y., ... Gutheil, J. C. (1995). An etoposide-resistant lung cancer subline overexpresses the multidrug resistance-associated protein. British Journal of Cancer, 72(3), 535-542. https://doi.org/10.1038/bjc.1995.370
Doyle, L. A. ; Ross, D. D. ; Ordonez, J. V. ; Yang, W. ; Gao, Y. ; Tong, Y. ; Belani, C. P. ; Gutheil, J. C. / An etoposide-resistant lung cancer subline overexpresses the multidrug resistance-associated protein. In: British Journal of Cancer. 1995 ; Vol. 72, No. 3. pp. 535-542.
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abstract = "We have characterised an etoposide-resistant subline of the small-cell lung cancer cell line, UMCC-1, derived at our centre. Subline UMCC-1/VP was developed by culturing the parent line in increasing concentrations of etoposide over 16 months. UMCC-1/VP is 20-fold resistant to etoposide by MTT assays, relative to the parent line, and is cross-resistant to doxorubicin, vincristine and actinomycin D, but not to taxol, cisplatin, melphalan, thiotepa or idarubicin. Topoisomerase II immunoblotting demonstrates a 50{\%} reduction of the protein in the resistant subline. The UMCC-1/VP subline demonstrates a marked decrease in the accumulation of [3H]etoposide relative to the parent line, as well as a modest reduction in the accumulation of daunorubicin. Reverse transcription-polymerase chain reaction assays demonstrate no detectable mdr1 expression but marked expression of the multidrug resistance-associated protein (MRP) gene in the resistant subline. Northern blotting with an MRP cDNA probe confirms marked overexpression of the MRP gene only in the UMCC-1/VP subline. Western blotting with antisera against MRP peptide confirms a 195 kDa protein band in the UMCC-1/VP subline. Southern blotting experiments demonstrate a 10-fold amplification of the MRP gene in the resistant subline. Depletion of glutathione with buthionine sulphoximine sensitised UMCC-1/VP cells to daunorubicin and etoposide. Our studies indicate that MRP gene expression may be induced by etoposide and may lead to reduced accumulation of the drug.",
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An etoposide-resistant lung cancer subline overexpresses the multidrug resistance-associated protein. / Doyle, L. A.; Ross, D. D.; Ordonez, J. V.; Yang, W.; Gao, Y.; Tong, Y.; Belani, C. P.; Gutheil, J. C.

In: British Journal of Cancer, Vol. 72, No. 3, 09.1995, p. 535-542.

Research output: Contribution to journalArticle

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T1 - An etoposide-resistant lung cancer subline overexpresses the multidrug resistance-associated protein

AU - Doyle, L. A.

AU - Ross, D. D.

AU - Ordonez, J. V.

AU - Yang, W.

AU - Gao, Y.

AU - Tong, Y.

AU - Belani, C. P.

AU - Gutheil, J. C.

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N2 - We have characterised an etoposide-resistant subline of the small-cell lung cancer cell line, UMCC-1, derived at our centre. Subline UMCC-1/VP was developed by culturing the parent line in increasing concentrations of etoposide over 16 months. UMCC-1/VP is 20-fold resistant to etoposide by MTT assays, relative to the parent line, and is cross-resistant to doxorubicin, vincristine and actinomycin D, but not to taxol, cisplatin, melphalan, thiotepa or idarubicin. Topoisomerase II immunoblotting demonstrates a 50% reduction of the protein in the resistant subline. The UMCC-1/VP subline demonstrates a marked decrease in the accumulation of [3H]etoposide relative to the parent line, as well as a modest reduction in the accumulation of daunorubicin. Reverse transcription-polymerase chain reaction assays demonstrate no detectable mdr1 expression but marked expression of the multidrug resistance-associated protein (MRP) gene in the resistant subline. Northern blotting with an MRP cDNA probe confirms marked overexpression of the MRP gene only in the UMCC-1/VP subline. Western blotting with antisera against MRP peptide confirms a 195 kDa protein band in the UMCC-1/VP subline. Southern blotting experiments demonstrate a 10-fold amplification of the MRP gene in the resistant subline. Depletion of glutathione with buthionine sulphoximine sensitised UMCC-1/VP cells to daunorubicin and etoposide. Our studies indicate that MRP gene expression may be induced by etoposide and may lead to reduced accumulation of the drug.

AB - We have characterised an etoposide-resistant subline of the small-cell lung cancer cell line, UMCC-1, derived at our centre. Subline UMCC-1/VP was developed by culturing the parent line in increasing concentrations of etoposide over 16 months. UMCC-1/VP is 20-fold resistant to etoposide by MTT assays, relative to the parent line, and is cross-resistant to doxorubicin, vincristine and actinomycin D, but not to taxol, cisplatin, melphalan, thiotepa or idarubicin. Topoisomerase II immunoblotting demonstrates a 50% reduction of the protein in the resistant subline. The UMCC-1/VP subline demonstrates a marked decrease in the accumulation of [3H]etoposide relative to the parent line, as well as a modest reduction in the accumulation of daunorubicin. Reverse transcription-polymerase chain reaction assays demonstrate no detectable mdr1 expression but marked expression of the multidrug resistance-associated protein (MRP) gene in the resistant subline. Northern blotting with an MRP cDNA probe confirms marked overexpression of the MRP gene only in the UMCC-1/VP subline. Western blotting with antisera against MRP peptide confirms a 195 kDa protein band in the UMCC-1/VP subline. Southern blotting experiments demonstrate a 10-fold amplification of the MRP gene in the resistant subline. Depletion of glutathione with buthionine sulphoximine sensitised UMCC-1/VP cells to daunorubicin and etoposide. Our studies indicate that MRP gene expression may be induced by etoposide and may lead to reduced accumulation of the drug.

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