umu.sePublications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
The platinum and copper metallome from sensitive and resistant T289 melanoma cells exposed to Cisplatin studied by a multi-analytical approach
Umeå University, Faculty of Science and Technology, Department of Chemistry.
Umeå University, Faculty of Medicine, Department of Surgical and Perioperative Sciences. Umeå University, Faculty of Medicine, Department of Public Health and Clinical Medicine, Medicine.
Umeå University, Faculty of Science and Technology, Department of Chemistry.
(English)Manuscript (preprint) (Other academic)
Abstract [en]

The copper and platinum metallome were evaluated in cisplatin (CDDP) sensitive and resistant melanoma T289 cells after exposure to cisplatin. Determination of the total concentrations of copper and platinum were combined with analysis of the intracellular copper and platinum profiles acquired by size exclusion chromatography (SEC) and reversed phase liquid chromatography (RPLC) coupled to inductively coupled plasma and electrospray ionization mass spectrometry (ICPMS and ESIMS, respectively). The total concentration of platinum in sensitive cells was 50-80% higher than in the resistant cell line and the intracellular copper concentration was decreased by ~30% when exposing the cells to cisplatin. SEC-ICPMS and RPLC-ICPMS profiles showed no obvious differences in the platinum profile between sensitive and resistant cells. However, the copper profiles differed showing a significant increase of some still unidentified copper species with an estimated molecular mass of 25-30 kDa. All together the data support the view that components of the copper homeostatic system are involved in CDDP accumulation and are affected by cisplatin exposure.

Keyword [en]
Metallome, HPLC-ICPMS, HPLC-ESIMS, Platinum, Copper
National Category
Analytical Chemistry
Research subject
Analytical Chemistry
Identifiers
URN: urn:nbn:se:umu:diva-37395OAI: oai:DiVA.org:umu-37395DiVA: diva2:360169
Available from: 2010-11-02 Created: 2010-11-02 Last updated: 2015-11-11Bibliographically approved
In thesis
1. Advances in analytical methodologies for studies of the platinum metallome in malignant cells exposed to cisplatin
Open this publication in new window or tab >>Advances in analytical methodologies for studies of the platinum metallome in malignant cells exposed to cisplatin
2010 (English)Doctoral thesis, comprehensive summary (Other academic)
Alternative title[sv]
Förbättrade analytiska metodologier för studier av platina-metallomet i maligna celler exponerade för cisplatin.
Abstract [en]

The scientific progress about the important chemotherapeutic drug substance cisplatin (CDDP) and its function has often been rendered by data difficult to interpret, and still many questions about its mode of action remains to be clarified by the scientific community. However, studies of CDDP possess a high complexity due to; i) low intracellular concentration, ii) many potential biomolecule targets, iii) poor or unknown stability of the intact drug and its biomolecule adducts and iv) complex and varying sample matrices. Metallomic studies, using advanced analytical techniques may contribute to clarify the interactions between CDDP and intracellular biomolecules. For a successful outcome sample preparation conditions as well as separation and detection techniques must be carefully selected and optimized to achieve accurate results and correct interpretation of data.

        This thesis describes some new and improved analytical methodologies for characterizing the Pt metallome in CDDP-exposed malignant cells. The developed methods are based on powerful liquid chromatography (LC) methods hyphenated to sensitive detection by inductively coupled plasma- (ICP) and electrospray ionization mass spectrometry (ESIMS). Consideration has also been taken about sample preparation conditions.

        By selecting “chemically inert” sample preparation (cell lysis by osmosis) and separation (using only nonreactive or no additatives) conditions we could avoid the formation of platinum artifact compounds previously described in the literature (Paper I and II). Using oxygen containing organic solvents with high boiling points (dimethylformamide; DMF, 1,4-dioxane, n-propanol and ethanol) as alternatives to acetonitrile in the LC separations, significant improvements were achieved in ICPMS sensitivity and robustness. When evaluated in combination with chromatographic performance and ESIMS detection the overall best performance was achieved with n-propanol (Paper II, III and IV). From the studies in Paper II we could show that free intact CDDP can be found in malignant cells, as supporting evidence for passive or endocytotic uptake of the drug and further estimate a half-life for intracellular CDDP to about 15 minutes. Such data has not been shown before. In Paper V, the above improved LC methods were used to demonstrate differences in the platinum and cupper metallome from sensitive and resistant T289 melanoma cells exposed to CDDP at near clinical levels.

        In a wider perspective we have shown the potential of using hydrophilic liquid interaction chromatography (HILIC) hyphenated to ICPMS detection as a general approach for analysis of hydrophilic metallo-compounds (Paper II). Taking advantage of the superior ICPMS performance using n-propanol gradients for reversed phase liquid chromatography (RPLC) possess a true alternative and /or complimentary technique to size exclusion chromatography (SEC) commonly applied within metallomic studies of biomolecules (Paper V). Using n-propanol in HILIC as well as in RPLC enables parallel detection by ICP- and ESIMS using only one set of chromatographic parameters (Paper III and IV), something commonly called for by scientists in the field.

Place, publisher, year, edition, pages
Umeå: Kemiska Institutionen, Umeå universitet, 2010. 8+44 p.
Keyword
Method development, Metallome, Inductively coupled plasma mass spectrometry, Electrospray ionization mass spectrometry, Liquid chromatography, Hyphenation, Organic modifier, Cisplatin, Platinum
National Category
Analytical Chemistry
Research subject
Analytical Chemistry
Identifiers
urn:nbn:se:umu:diva-37400 (URN)978-91-7459-085-2 (ISBN)
Public defence
2010-11-26, KBC-huset, Stora Hörsalen, KB3A1, Umeå Universitet, Umeå, 13:00 (English)
Opponent
Supervisors
Available from: 2010-11-05 Created: 2010-11-02 Last updated: 2010-11-05Bibliographically approved

Open Access in DiVA

No full text

Authority records BETA

Nygren, Yvonne

Search in DiVA

By author/editor
Nygren, Yvonne
By organisation
Department of ChemistryDepartment of Surgical and Perioperative SciencesMedicine
Analytical Chemistry

Search outside of DiVA

GoogleGoogle Scholar

urn-nbn

Altmetric score

urn-nbn
Total: 43 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf