Acquired Resistance to Peloruside A and Laulimalide is Associated with Downregulation of Vimentin in Human Ovarian Carcinoma Cells

作者：Kanakkanthara Arun; Rawson Pisana; Northcote Peter T; Miller John H^*

来源：Pharmaceutical Research, 2012, 29(11): 3022-3032.

DOI：10.1007/s11095-012-0773-x

• cancer resistance
• laulimalide
• ovarian carcinoma cells
• peloruside A
• vimentin

Acquired beta-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. %26lt;br%26gt;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. %26lt;br%26gt;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. %26lt;br%26gt;Downregulation of vimentin contributes to the resistance of 1A9 cells to the microtubule stabilizing agents, PLA and LAU.

• 出版日期2012-11