The roles of macromolecules in imatinib resistance of chronic myeloid leukemia cells by Fourier transform infrared spectroscopy

Baran, Yusuf; Ceylan, Cagatay; Camgöz, Aylin

doi:10.1016/j.biopha.2012.12.001

Cited by 13 publications

(17 citation statements)

References 45 publications

(57 reference statements)

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“…The effects of ponatinib on macromolecules in K562 and K562/IMA-3 cells We previously reported our FTIR spectroscopic comparison of imatinib-sensitive and imatinib-resistant K562 cells [20]. However, due to the changes induced by the resistance formation, the response of the drug-sensitive and drug-resistant cells to a second drug was not studied in terms of FTIR spectroscopic means.…”

Section: Resultsmentioning

confidence: 99%

“…This ratio is indicative of the RNA/DNA ratio [20] and was found to increase in the malignant cells when compared with the normal tissues.…”

Section: Discussionmentioning

confidence: 97%

“…The glucose/phosphate ratio is given as the intensity ratio of 1030 and 1086 cm −1 bands as a reliable measure of the metabolic turnover of the cells [41,20]. It was found to decrease by 4.94 % in the imatinib-sensitive and increase by 19.61 % in the imatinib-resistant K562 cells.…”

Section: Discussionmentioning

confidence: 99%

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A molecular and biophysical comparison of macromolecular changes in imatinib-sensitive and imatinib-resistant K562 cells exposed to ponatinib

et al. 2015

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Chronic myeloid leukemia (CML) is a type of hematological malignancy that is characterized by the generation of Philadelphia chromosome encoding BCR/ABL oncoprotein. Tyrosine kinase inhibitors (TKIs), imatinib, nilotinib, and dasatinib, are used for the frontline therapy of CML. Development of resistance against these TKIs in the patients bearing T315I mutation is a major obstacle in CML therapy. Ponatinib, the third-generation TKI, is novel drug that is effective even in CML patients with T315I mutation. The exact mechanism of ponatinib in CML has been still unknown. In this study, we aimed to determine the potential mechanisms and structural metabolic changes activated by ponatinib treatment in imatinib-sensitive K562 human CML cell lines and 3 μM-imatinib-resistant K562/IMA3 CML cell lines generated at our lab. Apoptotic and antiproliferative effects of ponatinib on imatinib-sensitive and 3 μM-imatinibresistant K562/IMA3 CML cells were determined by proliferation and apoptosis assays. Additionally, the effects of ponatinib on macromolecules and lipid profiles were also analyzed using Fourier transform infrared spectroscopy (FTIR). Our results revealed that ponatinib inhibited cell proliferation and induced apoptosis as determined by loss of mitochondrial membrane potential, increased caspase-3 enzyme activity, and transfer of phosphatidylserine to the plasma membrane in both K562 and K562/IMA-3 cells. Furthermore, cell cycle analyses revealed that ponatinib arrested K562 and K562/IMA-3 cells at G1 phase. Moreover, ponatinib treatment created a more ordered nucleic acid structure in the resistant cells. Although the lipid to protein ratio increased in imatinib-sensitive K562 cells with a little decrease in the K562/IMA-3 cells, ponatinib treatment indicated significant changes in the lipid composition such as a significant increase in the cellular cholesterol amounts much more in the K562/IMA-3 cells than the sensitive counterparts. Unsaturation in lipids was higher in the resistant cells; however, increases in lipids without phosphate and the number of acyl chains were much higher in the K562 cells. Taken together, all these results showed powerful antiproliferative and apoptotic effects of ponatinib in both imatinib-sensitive and imatinib-resistant CML cells in a dose-dependent manner, and hence, the use of ponatinib for the treatment of TKI-resistant CML patients may be an effective treatment approach in the clinic. More importantly, these results showed that FTIR spectroscopy can detect druginduced physiological changes in cancer drug resistance.

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Section: Resultsmentioning

confidence: 99%

“…This ratio is indicative of the RNA/DNA ratio [20] and was found to increase in the malignant cells when compared with the normal tissues.…”

Section: Discussionmentioning

confidence: 97%

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A molecular and biophysical comparison of macromolecular changes in imatinib-sensitive and imatinib-resistant K562 cells exposed to ponatinib

et al. 2015

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“…The trypsin spectrum was found to lose many secondary structural components as opposed to the original trypsin spectrum (Figure 5 b). The major sub bands that showed differences were that at 1694 cm -1 (1:antiparallel β-sheets), at 1658 cm -1 (2:α-helix), at 1637 cm -1 (3:β-sheets), and at 1627 cm -1 (4:aggregated β-sheets) [31].…”

Section: Trypsin Stability Studiesmentioning

confidence: 97%

Kinetic and structural characterization of interaction between trypsin and Equisetum arvense extract

Uslu¹,

Bayraktar²,

Ceylan³

2014

Turk J Bioch

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Kinetic and structural characterization of interaction between trypsin and Equisetum arvense extract [Trypsin ve Equisetum arvense özütü arasındaki etkileşimin kinetik ve yapısal karakterizasyonu] ABSTRACT Objective: In this study the inhibitory effect of E. arvense extract on trypsin activity and the effect of trypsin on E. arvense extract were studied. In addition the nature of the interaction between the extract and trypsin was investigated. Methods: The inhibitory effect ethanol extract of E. arvense on trypsin activity was determined using trypsin enzyme assay. The structural effects of the extract-trypsin interaction for the extract were analyzed by FTIR. Finally, the HPLC analyses were carried out to analyze the individual components of the extract and the supernatant and soluble precipitate phases. Results: E. arvense extract was found to decrease total percent activity of trypsin to 5% in 24 hour at 24 °C. FTIR analyses indicated that the interaction between trypsin and E. arvense extract caused changes in the structure and hydrogen bonding behavior and composition of the extract proteins. These interactions also caused the extract lipids to accumulate in the insoluble precipitate phase. Most of the phenolics remained in the supernatant phase enhancing the inactivation of trypsin. However, the precipitated compounds were shown to be of apolar in nature as shown in the HPLC chromatograms. Conclusion:The methods that were used showed that the high phenolic content of E. arvense was the main reason for the inhibition of trypsin enzyme activity by denaturing the enzyme. of 1:20. The dry leafs particles were shaked in the solution for overnight at 300 rpm at 24 °C. After the centrifugation at 2000xg for 5 minutes the supernatant was collected into 50 ml falcon tubes. The alcohol in the liquid phase was evaporated with a rotary evaporator. The aqueous extract was frozen for 24 hours and then freeze dried in a lyophilizer (Telstar-Cryodos). Preparation of inhibition solutionE. arvense extract was dissolved in distilled water to a final concentration of 10 mg/ml. By serial dilutions of stock solution 2 mg/ml, 0.2 mg/ml and 0.02 mg/ml concentrations were prepared. Preparation of trypsin solutionBovine pancreatic trypsin was purchased from Fluka Analytical (cat.93610) and used without further purification. Lyophilized powder of bovine pancreatic trypsin (Fluka Analytical, USA) was dissolved in 1 mM HCl to a final concentration of 0.1 mg/ml. Determination of enzyme kineticsTrypsin catalyzes the hydrolysis of N-benzoyl-L-arginine ethyl ester (BAEE) to N-benzoyl-L-arginine (BA) and ethanol. The slope of the absorbance versus time graph was the production rate of BA at pH 7.6.The reaction was held in a total volume of 3.2 ml quartz cuvette at 25 °C. The unit of trypsin activity was previously determined as 1 µmol BA produced per minute at 253 nm, at pH 7.6 and 25 °C. The reaction was carried out in 100 mM Tris buffer solution. To determine absorption of blank solution, 3050 μl tris buffer and 150 μl of BAEE solution ...

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“…In fact, the Philadelphia chromosome is characterized by its intrinsic hyper-mutability allowing the monitoring of kinase domain mutations most likely unfeasible 7 . In addition, there is high phenotypic variability within cells of a given tumour and leukaemic clones will contain sub-clones with different unpredictable grades of sensitivity/resistance to Vibrational microspectroscopy has been applied to study leukaemia cells' sensitivity/resistance to drugs in vitro [9][10][11][12][13][14][15][16][17][18][19][20][21] and to assess/monitor therapeutic responses in patients with leukaemia [22][23][24] . All these studies have proven that vibrational spectroscopy, performed either with FTIR or Raman techniques, is sensitive enough to identify some biochemical signatures related to the effects of different anti-leukemic drugs.…”

Section: Introductionmentioning

confidence: 99%

Effects of nilotinib on leukaemia cells using vibrational microspectroscopy and cell cloning

Siddique

Rutter

Wehbe

et al. 2017

Analyst

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Over the last few years, both synchrotron-based FTIR (S-FTIR) and Raman microspectroscopies have helped to better understand the effects of drugs on cancer cells. However, cancer is a mixture of cells with different sensitivity/resistance to drugs. Furthermore, the effects of drugs on cells produce both chemical and morphological changes, the latter could affect the spectra of cells incubated with drugs. Here, we successfully cloned sensitive and resistant leukaemia cells to nilotinib, a drug used in the management of leukaemia. This allowed both the study of a more uniform population and the study of sensitive and resistant cells prior to the addition of the drug with both S-FTIR and Raman microspectroscopies. The incubation with nilotinib produced changes in the S-FTIR and Raman spectra of both sensitive and resistant clones to nilotinib. Principal Component Analysis was able to distinguish between cells incubated in the absence or presence of the drug, even in the case of resistant clones. The latter would confirm that the spectral differences between the so-called resistant clonal cells prior to and after adding a drug might reside on those more or less sensitive cells that have been able to remain alive when they were collected to be studied with S-FTIR or Raman microspectroscopies. The data presented here indicate that the methodology of cell cloning can be applied to different types of malignant cells. This should facilitate the identification of spectral biomarkers of sensitivity/resistance to drugs. The next step would be a better assessment of sensitivity/resistance of leukaemia cells from patients which could guide clinicians to better tailor treatments to each individual patient.3

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The roles of macromolecules in imatinib resistance of chronic myeloid leukemia cells by Fourier transform infrared spectroscopy

Cited by 13 publications

References 45 publications

A molecular and biophysical comparison of macromolecular changes in imatinib-sensitive and imatinib-resistant K562 cells exposed to ponatinib

A molecular and biophysical comparison of macromolecular changes in imatinib-sensitive and imatinib-resistant K562 cells exposed to ponatinib

Kinetic and structural characterization of interaction between trypsin and Equisetum arvense extract

Effects of nilotinib on leukaemia cells using vibrational microspectroscopy and cell cloning

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