Acquired resistance to peloruside A and laulimalide is associated with downregulation of vimentin in human ovarian carcinoma cells.

Abstract

PURPOSE:

Acquired β-tubulin alterations in human ovarian carcinoma 1A9 cells were previously shown to confer resistance to the microtubule stabilizing agents peloruside A (PLA) and laulimalide (LAU). We examined the proteome of resistant cells to see what other protein changes occurred as a result of the acquired drug resistance.

METHODS:

Two-dimensional differential in-gel electrophoresis was performed to explore differentially expressed proteins in the resistant 1A9-R1 (R1) and 1A9-L4 (L4) cells. The proteins on the gels were identified by MALDI-TOF MS, and altered protein abundance was confirmed by Western blotting and immunocytochemistry. Vimentin expression was restored in vimentin-deficient L4 cells by transfecting a full-length human vimentin cDNA, and sensitivity to PLA and LAU were tested using an MTT cell proliferation assay.

RESULTS:

Proteomic analysis identified several proteins that were significantly altered in the resistant cells relative to the parental 1A9 cells. Using Western blotting and immunocytochemistry, a decreased vimentin abundance in the L4 cells was validated. Vimentin levels were unchanged in PLA-resistant R1 cells and paclitaxel/epothilone-resistant derivatives of 1A9 cells. Vimentin cDNA transfection into L4 cells partially restored PLA and LAU sensitivity.

CONCLUSIONS:

Downregulation of vimentin contributes to the resistance of 1A9 cells to the microtubule stabilizing agents, PLA and LAU.

• Authors: Arun Kanakkanthara, Pisana Rawson, Peter T Northcote, John H Miller
• Journal: Pharmaceutical research (Pharm Res) Vol. 29 Issue 11 Pg. 3022-32 (Nov 2012) ISSN: 1573-904X [Electronic] United States
• PMID: 22584948 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antineoplastic Agents
• Bridged Bicyclo Compounds, Heterocyclic
• DNA, Complementary
• Lactones
• Macrolides
• Vimentin
• laulimalide
• peloruside A

Topics

• Antineoplastic Agents (pharmacology)
• Bridged Bicyclo Compounds, Heterocyclic (pharmacology)
• Carcinoma (drug therapy, genetics, metabolism)
• Cell Line, Tumor
• DNA, Complementary (genetics)
• Down-Regulation (drug effects)
• Drug Resistance, Neoplasm
• Female
• Gene Expression Regulation, Neoplastic (drug effects)
• Humans
• Lactones (pharmacology)
• Macrolides (pharmacology)
• Microtubules (drug effects, genetics, metabolism)
• Ovarian Neoplasms (drug therapy, genetics, metabolism)
• Proteomics (methods)
• Transfection
• Vimentin (biosynthesis, genetics, metabolism)