MicroRNA 146-5p, miR-let-7c-5p, miR-221 and miR-345-5p are differentially expressed in Respiratory Syncytial Virus (RSV) persistently infected HEp-2 cells.

Many viruses can establish non-cytolytic, chronic infections in host cells. Beyond the intrinsically interesting questions of how this long-term parasitism is achieved, persistently infected cells can be useful to study virus-host interactions. MicroRNAs (miRNAs) are a class of noncoding RNAs transcribed from the genomes of all multicellular organisms and some viruses. Individual miRNAs may regulate several hundred genes. In this research we have studied the expression of a selective group of host-cell encoded miRNAs, as expressed in a Respiratory Syncytial Virus (RSV) persistently infected HEp-2 cell line (HEp-2 + RSV-GFP). The RSV is a virus that does not encode miRNAs in its genome. Our study shows that Dicer is down regulated, miRNA's 146a-5p is strongly up-regulated and miRNAs 345-5p, let-7c-5p and miRNA's-221 are down-regulated in HEp-2 + RSV-GFP cells. Correspondingly, changes in the miRNA 146a-5p and he sequences of the reference genes are miRNA 345-5p respective miRNAs target proteins: HSP-70 and p21, were observed. Thus, RSV persistent viral infection induces unique patterns of miRNA's expression with relevance to how the virus regulates the host cell response to infection.

Grading of intrinsic and acquired cisplatin-resistant human melanoma cell lines: an infrared ATR study.

Attenuated total reflection (ATR) spectroscopy is used as an in vitro optical approach for the diagnosis and characterization of cell and tissue pathology. In comparison with the more conventional FTIR microspectroscopy that relies on transmission of IR radiation, ATR spectroscopy uses the evanescent wave technique, which is a step forward toward in vivo research. The aim of the present investigation was to examine the potential of ATR spectroscopy to differentiate between drug-resistant and drug-sensitive melanoma cell lines. We studied two human melanoma parental cell lines, GA and BG, and their cisplatin-resistant counterparts, GAC and BGC, respectively, which were derived by survival selection with this anticancer drug. Cisplatin cytotoxicity was measured on the four cell lines, and their relative resistance to cisplatin was established: BGC > BG > GAC > GA. Different resistance mechanisms were noticed between the two parental groups in accordance with their spectrum. ATR spectra-based cluster analysis of the selective biomarkers, such as phosphate and RNA/DNA, were found useful in differentiating sensitive from resistant cells. Normalized and absolute values of the differences between spectra were employed to compare between the two parental groups. It was possible to predict the relative cisplatin resistance between the cell lines using the discriminant classifying function. The success rates in predicting cisplatin resistance in these cells was 88 and 81% for GA versus GAC and BG versus BGC, respectively. These results support the further development of the ATR technique as a simple, in vitro, reagent-free method to identify drug resistance in cancer cells.

Discrimination between drug-resistant and non-resistant human melanoma cell lines by FTIR spectroscopy.

We investigated the ability of FTIR-microscopy to define spectral changes between drug-sensitive and drug-resistant human melanoma cells. As a model system, a resistant melanoma cell line (GAC) was selected with cisplatin from parental (GA) cells. Using Fourier transform infrared spectroscopy (FTIR) we investigated the ability to differentiate between the resistant variant derived from the sensitive parental cell line, in the absence of cisplatin. We determined and validated spectral parameters (biomarkers) that differentiated between the two cell lines. By applying the principal component analysis (PCA) model, we reduced the original data size to six principal components. We detected a significant and consistent increase in the cell's DNA/RNA ratio as well as an increase in the lipid/protein ratio in the resistant cells. These results strongly support the potential of developing FTIR microspectroscopy as a simple, reagent-free method for the identification of drug-resistant cells. Rapid detection of tumors resistant to a particular drug, should contribute to the ability of the physician to choose an effective treatment protocol.

Effects of hyperthermia and drug in cultured mammalian cells.

Hyperthermia (42 degrees C) and peptichemio (PTC) applied simultaneously result in an increased killing effect and in a significant induction of single strand breaks (SSB) of DNA molecule. The enhanced killing effect was observed in all circumstances, whenever hyperthermia was applied before, during or after PTC treatment, but the most effective cell killing was obtained when hyperthermia and PTC were applied simultaneously. The results show that PTC concentrations used in these experiments do not induce SSB in DNA molecule during incubation for 2 hours at 37 degrees C. When the cells were exposed to PTC and hyperthermia the induction of SSB was observed. The number of SSB depends on the exposure time and PTC concentration. Cells with a greater number of SSB lost the capacity for repair during post-incubation at 42 degrees C.