Analysis of the influence of adalimumab to the expression pattern of mRNA and protein of TGF-ß1-3 in dermal fibroblast exposed to lipopolysaccharide.

INTRODUCTION: Psoriasis is a inflammatory illness, where incorrect expression of cytokines and bacteria lipopolysaccharide are observed. In the therapy of moderate to severe psoriasis anti-TNF drugs, i.e. adalimumab are used which have the influence for secreting another cytokines, such as transforming growth factor-ß (TGF-ß). AIM: To analyse the expression profile of mRNA TGF-ß1-3 and proteins (TGF-ß1 and TGF-ß2) it codes in normal human dermal fibroblasts (NHDF) exposed to bacterial lipopolysaccharide (induction of inflammation) and adalimumab (anti-TNF drug). MATERIAL AND METHODS: NHDFs treated with bacterial lipopolysaccharide at a medium concentration of 1 µg/ml for 8 h, and then added to an adalimumab culture at a concentration of 8 µg/ml and continued exposure of the fibroblasts to it for 2, 8 and 24 h. The molecular analysis included microarray, RTqPCR and ELISA assays. RESULTS: Treating the skin fibroblast cells with LPS resulted in significant statistical changes in the expression of TGF-ß1 (↑) and TGF-ß2 (↓) in comparison to the control culture. Likewise, after adding adalimumab to the culture of NHDF treated previously with LPS, significant changes in the expression of TGF-ß1 (↑) and TGF-ß2 (↓) were noted in comparison to the control culture (p < 0.05). On the protein level it can be determined that LPS and adalimumab cause an increase in the concentration of TGF-ß1 and a decrease in the expression of TGF-ß2 in comparison to the control culture. CONCLUSIONS: Blocking the signalling dependant on TNF-α using adalimumab causes an increase in the expression of TGF-ß1 and a simultaneous decrease in the case of TGF-ß2.

Effect of adalimumab on the expression profile of mRNA, and protein associated with JAK/STAT signaling pathway in fibroblast exposed to lipopolysaccharide.

The aim of this study was to assess the effect of adalimumab on the expression level of mRNA and protein TNF-α, IFN-γ, IL-17, IL12A, IL12B, and IL23A in the culture of normal human fibroblasts, in which the LPS inflammation process was induced. The NHDF culture was exposed to the effect of LPS in the concentrations of 1, 2, and 10 µg/mL for 2, 8, and 24 hour periods, and then adalimumab was added at the concentration of 8 µg/mL, it was then incubated for 2, 8, and 24 hour. Cells unexposed to LPS and adalimumab constituted the control. The microarray expression techniques, RTqPCR, and ELISA assay were used. Irrespectively of the concentration of LPS used and the incubation time of it with cells overexpression of the analyzed genes is present, with increasing factor concentration used to induce inflammation and incubation time with it, expression of the assessed genes was greater. In turn, adding the anti-TNF drug to the culture caused the silencing of the expression of the mRNAs and proteins. It was confirmed that LPS and adalimumab above all affect the expression of genes and proteins dependent on the interaction of IL-12 with receptors, which are TNF-α and IFN-γ, and to a lesser extent also modulate IL-17.

Variances in the Level of COX-2 and iNOS in Different Grades of Endometrial Cancer.

BACKGROUND: Many experimental studies have demonstrated the importance of COX-2 in the tumor angiogenesis. Inducible iNOS is responsible for a high and stable level of nitric oxide and is expressed in response to pro-inflammatory factors. OBJECTIVE: The aim of this study was to evaluate the expression of COX-2 and iNOS at the protein level and to assess their potential prognostic significance in patients with endometrial cancer. METHODS: The study group consisted of 45 women with endometrial cancer divided according to the degree of histological differentiation i.e. G1, 17; G2, 15; G3, 13. The control group consisted of 15 women without neoplastic changes. The expression of studied proteins was determined immunohistochemically with specific polyclonal antibodies. RESULTS: Analysis of the COX-2 expression showed that the optical density of the reaction product in G1 reached 186% in the control group, while the values in G2 and G3 reached 243% and 293%, respectively. In the case of iNOS, the optical density of the reaction product reached the following percentages in the control group: 147% in G1, 243% in G2, and 241% in G3. CONCLUSION: Our findings suggest that changes in the expression of COX-2 and iNOS may be potentially useful in predicting the progression of endometrial cancer and treatment effectiveness.