Abstract

The aim of this project is to use electron probe x-ray microanalysis to determine the mechanism for entry of the anti-cancer drug cis-diamminedichloroplatinum II (cisplatin) into cancer cells, and how this entry is affected by drug resistance. Drug-sensitive and drug-resistant liver carcinoma cells were incubated for different times (0.5 hr, 1 hr, 2 hr, 4 hr) with 400 micromolar cisplatin and 100 micrograms/ml ferric chloride. The enzyme trypsin was used to release the cells gently from the culture dishes and to avoid damage by scraping. Cells were then pelleted, rapidly frozen in liquid ethane and cryosectioned. In the absence of cisplatin, cells prepared in this way were found to have normal Na/K gradients across the plasma membrane. Experiments are being performed on treated cells using x-ray microanalysis to co-localize cisplatin and iron.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
Office of The Director, National Institutes of Health (OD)
Type
Intramural Research (Z01)
Project #
1Z01OD010491-03
Application #
6432968
Study Section
(BEPS)
Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
2000
Total Cost
Indirect Cost

  Institution

Name
Office of the Director, NIH
Department
Type
DUNS #
City
State
Country
United States
Zip Code

  Publications

Licht, T; Goldenberg, S K; Vieira, W D et al. (2000) Drug selection of MDR1-transduced hematopoietic cells ex vivo increases transgene expression and chemoresistance in reconstituted bone marrow in mice. Gene Ther 7:348-58
Lee, C G; Ramachandra, M; Jeang, K T et al. (2000) Effect of ABC transporters on HIV-1 infection: inhibition of virus production by the MDR1 transporter. FASEB J 14:516-22
Shen, D W; Goldenberg, S; Pastan, I et al. (2000) Decreased accumulation of [14C]carboplatin in human cisplatin-resistant cells results from reduced energy-dependent uptake. J Cell Physiol 183:108-16
Hafkemeyer, P; Licht, T; Pastan, I et al. (2000) Chemoprotection of hematopoietic cells by a mutant P-glycoprotein resistant to a potent chemosensitizer of multidrug-resistant cancers. Hum Gene Ther 11:555-65
Booth, C L; Pulaski, L; Gottesman, M M et al. (2000) Analysis of the properties of the N-terminal nucleotide-binding domain of human P-glycoprotein. Biochemistry 39:5518-26
Cowan, K H; Moscow, J A; Huang, H et al. (1999) Paclitaxel chemotherapy after autologous stem-cell transplantation and engraftment of hematopoietic cells transduced with a retrovirus containing the multidrug resistance complementary DNA (MDR1) in metastatic breast cancer patients. Clin Cancer Res 5:1619-28
Zhou, Y; Gottesman, M M; Pastan, I (1999) Domain exchangeability between the multidrug transporter (MDR1) and phosphatidylcholine flippase (MDR2). Mol Pharmacol 56:997-1004
Ambudkar, S V; Dey, S; Hrycyna, C A et al. (1999) Biochemical, cellular, and pharmacological aspects of the multidrug transporter. Annu Rev Pharmacol Toxicol 39:361-98
Zhou, Y; Gottesman, M M; Pastan, I (1999) Studies of human MDR1-MDR2 chimeras demonstrate the functional exchangeability of a major transmembrane segment of the multidrug transporter and phosphatidylcholine flippase. Mol Cell Biol 19:1450-9
Dey, S; Hafkemeyer, P; Pastan, I et al. (1999) A single amino acid residue contributes to distinct mechanisms of inhibition of the human multidrug transporter by stereoisomers of the dopamine receptor antagonist flupentixol. Biochemistry 38:6630-9

Showing the most recent 10 out of 16 publications