Inhibition of farnesyl protein transferase sensitizes human MCF-7 breast cancer cells to roscovitine-mediated cell cycle arrest.
We reported recently that roscovitine (ROSC), a selective cyclin-dependent kinase (CDK) inhibitor, arrests human MCF-7 breast cancer cells in G(2) phase of the cell cycle, and concomitantly induces apoptosis. Human MCF-7 breast cancer cells are known to express elevated levels of c-Ha-Ras protein. To achieve full biological activity, de novo synthesized c-Ha-Ras protein has to be farnesylated and after further processing it needs to be attached to the plasma membrane. Therefore, we decided to prove whether prevention of protein farnesylation would sensitize MCF-7 cells to the action of ROSC. MCF-7 cells were treated with 1-40 microM ROSC alone, or in combination with L-744,832, an inhibitor of farnesyl protein transferase (FTPase). To measure the impact on the proliferation of the cells, we used the CellTiterGlo viability assay and FACS analysis was employed to quantify the DNA-content of the single cells. The amount and phosphorylation status of relevant proteins after lysis of MCF-7 cells was assessed on Western blots using (phospho)-specific antibodies. The combined treatment with L-744,832 and ROSC for 24 h, markedly reduced the number of viable MCF-7 cells, primarily, by re-enforcing the cell cycle arrest. Interestingly, the potentiation of the ROSC-mediated inhibition of cell proliferation became evident during the 48 h post-incubation period in presence of the FPTase inhibitor. Inhibition of FPTase in ROSC-treated cells reduced the number of viable cells by approximately 30%. Evidently, the combined treatment sensitizes MCF-7 cells to the action of ROSC, thereby allowing to reduce the dose of the drug and to minimize side effects.
Wesierska-Gadek
Józefa
J
Department of Medicine I, Div.: Institute of Cancer Research, Vienna Medical University, Vienna, Austria. Jozefa.Gadek-Wesierski@meduniwien.ac.at
Maurer
Margarita
M
Schmid
Gerald
G
D002453:Cell Cycle; D045744:Cell Line, Tumor; D002469:Cell Separation; D051231:Farnesyltranstransferase / Q000037:antagonists & inhibitors*; D005434:Flow Cytometry; D016195:G2 Phase; D016158:Genes, p53; D006801:Humans; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism; D011687:Purines / Q000494:pharmacology*; D000077546:Roscovitine; D015398:Signal Transduction; D013347:Subcellular Fractions; D013997:Time Factors; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D047428:Protein Kinase Inhibitors; D011687:Purines; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D000077546:Roscovitine; D051231:Farnesyltranstransferase; D016283:Proto-Oncogene Proteins p21(ras)