SIDE EFFECTS OF SUNITINIB MANIFESTED ON SKIN OF PATIENTS DIAGNOSED WITH RENAL CELL CARCINOMA: CASE-CONTROL STUDY

Sunitinib is a small, lipophilic, synthetic molecule that interferes with tyrosinekinase domain of vascular endothelial growth factor receptor, and prevents its activation after binding the vascular endothelial growth factor. Both in vitro and in vivo, sunitinib inhibits angiogenesis, and suppress growth of metastases, which depends on newly formed blood vessels. It was approved by FDA (U.S. Food and Drug Administration) in January 2006 for the treatment of advanced renal-cell cancer and imatinib-resistant gastrointestinal stromal tumours.Among other adverse effects, it causes skin desquamation on fingers and toes, whitening of hair, eyebrows, mustache and beard. The aim of our study was to prove the causal relationship between sunitinib administration and skin adverse effects.The study involved the patients with metastasized renal cell carcinoma treated at the Institute for Radiology and Oncology of Serbia, in Belgrade. There were twelve patients (mean age 53.3 ± 11.1 years) who took sunitinib 50 mg daily, for 4 weeks (“cases”). Control group was composed of fourteen patients (mean age 54.6 ± 9.8 years) on standard therapy with interferon alpha (6 MJ three times weekly) and vinblastine 10mg, two days per cycle. The control patients were matched with “cases” by age, sex, phase of the disease and nephrectomy.Out of the patients who received sunitinib, eleven (92%) patients developed desquamation on fingers and toes, whitening of hair, eyebrows, mustache and beard, while none of the skin adverse effects was observed in the control group (OR = 143). The distribution of hypertension, heart diseases and diabetes mellitus in the groups was not significantly different (p>0.05).There is a strong association between administration of sunitinib in patients with renal cell carcinoma and observed skin adverse effects

Detection of DNA mutations based on analysis of multiple wavelength excitation/emission fluorescence kinetics curves in real-time PCR

The key method for therapies of various cancer types could be the molecular-targeted therapy, based on individual gene profile for each patient. One of the main procedures used for genetic testing is the real-time polymerase chain reaction (real-time PCR). Physical principle behind real-time PCR procedure is the fluorescence. Fluorescence labeled probes (primers) is attached to quenchers. Upon reaction of polymerization, quenchers are removed, and the fluorescence emission intensity increases in time. Emission spectra shape and its maximum position can differ if the fluorophore was present in different microenvironment. That property is widely exploited in fluorescence spectroscopy and chromatography. This paper, for the first time, describes utilization of full spectroscopic potential of multichannel excitation/emission filter sets in real-time PCR device. Instead of monitoring fluorescence intensity in time for a single fluorescence emission channel, the ratio values of three different kinetics curves were calculated and analyzed by applying k-means clustering and dendrogram analysis. Obtained results have shown that described analytical improvement provides identification of nine different groups of mutations if the commercial QIAGEN (R) EGFR PCR Kit was used. Method can be applied to any kit, capable to simultaneously detect several different mutations

Detection of DNA mutations based on analysis of multiple wavelength excitation/emission fluorescence kinetics curves in real-time PCR

The key method for therapies of various cancer types could be the molecular-targeted therapy, based on individual gene profile for each patient. One of the main procedures used for genetic testing is the real-time polymerase chain reaction (real-time PCR). Physical principle behind real-time PCR procedure is the fluorescence. Fluorescence labeled probes (primers) is attached to quenchers. Upon reaction of polymerization, quenchers are removed, and the fluorescence emission intensity increases in time. Emission spectra shape and its maximum position can differ if the fluorophore was present in different microenvironment. That property is widely exploited in fluorescence spectroscopy and chromatography. This paper, for the first time, describes utilization of full spectroscopic potential of multichannel excitation/emission filter sets in real-time PCR device. Instead of monitoring fluorescence intensity in time for a single fluorescence emission channel, the ratio values of three different kinetics curves were calculated and analyzed by applying k-means clustering and dendrogram analysis. Obtained results have shown that described analytical improvement provides identification of nine different groups of mutations if the commercial QIAGEN (R) EGFR PCR Kit was used. Method can be applied to any kit, capable to simultaneously detect several different mutations

Cytotoxic activity of ethanol extracts of in vitro grown Cistus creticus subsp creticus L. on human cancer cell lines

Cistus creticus subsp. creticus is a native plant of the Mediterranean region and it has been used since ancient times for its medicinal properties. The leaves secrete labdane type diterpenes that exhibit considerable antibacterial and cytotoxic activities. However there are no data about implementation of in vitro culture of C. creticus in order to obtain applicable production of labdane diterpenes. We successfully established in vitro culture of shoots and roots on solid or liquid MS (Murashige and Skoog) medium without addition of plant growth regulators. GC/FID and GC/MS detections were used to carry out characterization of ethanol extracts of shoots and roots from in vitro culture. Labdane diterpenes were the most abundant compounds in shoot extract, but absent from the root extract. Total phenolic and flavonoid contents of C. cretins shoot and root ethanol extracts were also determined. In order to evaluate possible cytotoxic activity of extracts, sulforhodamine B (SRB) assay was performed on five human cancer cell lines. Only shoot extract had cytotoxic activity on HeLa (cervix), MDA-MB-453 (breast) and FemX (melanoma) cancer cells with IC50 reaching 80.83 mu g/ml (HeLa), 76.18 mu g/ml (MDA-MB-453), and 87.52 mu g/ml (FemX) respectively. This is the first report of cytotoxic effect of labdane diterpenes from Cistus on HeLa cells. (C) 2012 Elsevier B.V. All rights reserved.Ministry of Education and Science of the Republic of Serbia [173024, III41026, 173021