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Published on: January 21, 2015
Macromolecular changes in nilotinib resistant K562 cells; an in vitro study by Fourier transform infrared
Cagatay Ceylan1, Aylin Camgoz, Yusuf Baran
1Izmir Institute of Technology, Department of Food Engineering, Urla, Izmir, Turkey. cagatayceylan@iyte.edu.tr
Abstract:
Nilotinib is a second generation tyrosine kinase inhibitor which is used in both first and second line treatment of chronic myeloid leukemia (CML). In the present work, the effects of nilotinib resistance on K562 cells were investigated at the molecular level using Fourier transform infrared (FT-IR) spectroscopy. Human K562 CML cells were exposed to step-wise increasing concentrations of nilotinib, and sub-clones of K562 cells resistant to 50nM nilotinib were generated and referred to as K562/NIL-50 cells. Antiproliferative effects of nilotinib were determined by XTT cell proliferation assay. Changes in macromolecules in parental and resistant cells were studied by FT-IR spectroscopy. Nilotinib resistance caused significant changes which indicated increases in the level of glycogen and membrane/lipid order. The amount of unsaturated lipids increased in the nilotinib resistant cells indicating lipid peroxidation. The total amount of lipids did not change significantly but the relative proportion of cholesterol and triglycerides altered considerably. Moreover, the transcriptional status decreased but metabolic turn-over increased as revealed by the FT-IR spectra. In addition, changes in the proteome and structural changes in both proteins and the nucleus were observed in the K562/NIL-50 cells. Protein secondary structural analyses revealed that alpha helix structure and random coil structure decreased, however, anti-parallel beta sheet structure, beta sheet structure and turns structure increased. These results indicate that the FT-IR technique provides a method for analyzing drug resistance related structural changes in leukemia and other cancer types.
Insights
Nilotinib resistance in chronic myeloid leukemia (CML) cells alters cellular macromolecules. Fourier transform infrared spectroscopy revealed significant changes in lipids, glycogen, and protein structures, offering insights into drug resistance mechanisms.
Area of Science:
- Biochemistry
- Molecular Biology
- Spectroscopy
Background:
- Nilotinib is a tyrosine kinase inhibitor crucial for chronic myeloid leukemia (CML) treatment.
- Understanding nilotinib resistance mechanisms at the molecular level is vital for improving CML therapy.
- K562 cells are a human leukemia cell line commonly used in CML research.
Purpose of the Study:
- To investigate the molecular effects of nilotinib resistance on K562 CML cells.
- To analyze changes in cellular macromolecules using Fourier transform infrared (FT-IR) spectroscopy.
- To identify potential biomarkers for nilotinib resistance.
Main Methods:
- Generation of nilotinib-resistant K562 cells (K562/NIL-50) through stepwise drug exposure.
- XTT cell proliferation assay to assess antiproliferative effects.
- Fourier transform infrared (FT-IR) spectroscopy to analyze macromolecular composition and structure.
Main Results:
- Nilotinib resistance led to increased glycogen levels and enhanced membrane/lipid order.
- Increased unsaturated lipids indicated lipid peroxidation in resistant cells.
- Significant alterations in cholesterol and triglyceride proportions were observed.
- FT-IR spectra revealed decreased transcriptional status but increased metabolic turnover.
- Changes in proteome and structural modifications in proteins and the nucleus were detected.
- Protein secondary structure analysis showed decreased alpha helix and random coil, with increased beta sheet and turn structures.
Conclusions:
- FT-IR spectroscopy is a valuable tool for analyzing drug resistance-related structural changes in leukemia.
- Nilotinib resistance is associated with significant alterations in cellular lipid and protein metabolism and structure.
- These findings contribute to a deeper understanding of CML drug resistance mechanisms.
