The effect of cisplatin on human larynx carcinoma cell motility.

Head and neck tumors are one of the major public health problem all over the world. Cellular response of larynx carcinoma to cisplatin (CDDP) chemotherapy occurs both in cell-interdependent and cell-autonomous pathways. In the first pathway, cytotoxic signal transduction is mediated via gap-junctional intercellular communication (GIJC). CDDP also influence tumor cell migration.The aim of this study was the analysis of the effect of CDDP (0.5 microg/ml and 1.5 microg/ml) on the gap-junction intercellular communication and motility, respectively, in two new cell cultures (RK33 and RK45) derived from human larynx carcinoma. The migration of RK45 cell line was slightly inhibited and RK33 not affected after the incubation with CDDP. Tumor cells incubation with CDDP resulted in farther LY migration through neighboring cells beyond monolayer wound than in control cultures.In conclusion, there is a relationship between intercellular communication via gap junctions and motility of laryngeal tumor cells after CDDP application

The impact of oleanolic and ursolic acid on corneal epithelial cells in vitro

INTRODUCTION. Oleanolic (OA) and ursolic (UA) acids belong to the triterpene group widely present in plants. These compounds are recognised to have anti-inflammatory properties and thus are considered to be used in therapies as well as in cosmetic, natural health, or diet products. AIM. The scientific hypothesis of our study was to show that OA and UA influence corneal epithelial cells cultured in vitro. METHODS. Toxicity tests, based on MTT and Neutral Red (NR) uptake, measurement of nitric oxide (NOx) level, as well as analysis of metalloproteinases (MMP-2 and MMP-9) amount and activity were performed. RESULTS . UA expressed significantly higher toxicity on cells than OA. At the lowest concentration applied (5 μM), UA limited cellular metabolism and viability on average by 22% as compared to untreated control, while 25 μM resulted in values lower than 10%. On the other hand, OA at the highest (100 μM) concentration limited cellular metabolism and viability by about 20%. NOx level significantly increased when OA and UA were applied at concentrations of 25 and 100 μM, respectively. OA and UA had a stronger impact on the level of MMP-2 than MMP-9. OA and UA reduced MMP-2 and MMP-9 in the whole range of concentrations. Tested triterpenoids had no significant impact on MMP activity. CONCLUSIONS. OA and UA have a different impact on human corneal epithelial cells. UA is toxic for corneal epithelial cells, while OA exhibits milder activity, which may be useful for further analysis in ocular pharmacology

Reactivity of corneal and conjunctival epithelial cells to lipopolysaccharide (LPS) and/or irradiation with visible light in vitro

INTRODUCTION. Visible light and inflammation caused by bacterial endotoxins strongly influence direct cell interactions and modulate the expression of selected factors, such as nitric oxide (NO) and cyclooxygenase-2 (COX-2). The aim of the study is to establish whether exposition of corneal or conjunctival epithelial cells to visible light and/ or LPS may change their viability, direct cellular interactions and expression of NO and COX-2. MATERIALS AND METHODS. In vitro cultured human corneal and conjunctival epithelial cells were used in the study. The following assays were performed: Neutral Red (NR) uptake, nitric oxide (NO) quantification by the Griess method, cytoskeletal F-actin organization by fluorescent staining, and COX-2 expression by immunofluorescence. RESULTS. LPS reduced the viability of the cells, especially conjunctival epithelial cells. All cell stimulation variants tested (visible light and/or LPS treatment) led to decreased nitric oxide (NO) production both by corneal and conjunctival epithelial cells. No changes in cytoskeletal F-actin filaments were observed after the cells had been treated with light or the endotoxin. LPS slightly increased COX-2 expression, but light had no, or a slightly reducing, effect on the level of this enzyme. CONCLUSIONS. Visible light and/or bacterial endotoxin (LPS) may, depending on the local microenvironmental conditions, cooperate or interfere with each other’s activity in inducing ocular surface inflammation

The effect of ursolic and oleanolic acids on human skin fibroblast cells

In this article, we look at how ursolic and oleanolic acids can be used for the purpose of quality control of natural products used in dermatocosmetology as well as of various other therapeutic preparations. Ursolic acid (UA) and oleanolic acid (OA) are pentacyclic triterpenes and they are constituents of many medicinal herbs. In this study, we analyzed the cytotoxic and anti-proliferative activity of OA and UA against normal human skin fibroblasts (HSF). Additionally, the scavenging activity of free radicals of both acids was analyzed. The sensitivity of cells to OA and UA activity was determined using a standard spectrophotometric (MTT) assay. The free radical scavenging activity of OA and UA was measured using the DPPH• test. The F-actin cytoskeletal proteins organization was analyzed using TRITC-phalloidine fluorescent staining. The cytotoxic activity of the analyzed acids was determined using Neutral Red (NR) uptake assay. Of the two isomeric compounds, UA showed a higher cytotoxic activity against HSF cells than did OA. Our investigations showed that OA, in view of its non-toxic nature, may be used as a supplementary factor for dermal preparations. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 4, pp. 664–669

Cell death in HeLa cells upon imperatorin and cisplatin treatment

There is growing evidence that commonly applied chemotherapy regimens can be improved by introducingnew, specific, active and low side-effect drugs, or by combining substances to obtain the required clinicaleffect. The aim of the present study was to investigate the effects of imperatorin and cisplatin, applied separatelyor in combination, on apoptosis, necrosis and autophagy induction in the human cervical carcinoma cell line(HeLa). Imperatorin appeared to be a potent autophagy inducer, rather than a necrotic or apoptotic one. Incontrast, cisplatin induced mainly apoptosis and necrosis after 6 h and 24 h, while longer incubation resultedonly in necrosis induction. When HeLa cells were incubated with both drugs, autophagy appeared most frequently,although to a smaller extent than that observed after imperatorin administered alone. At the molecularlevel, autophagy was correlated with the presence of the cleaved form of microtubule-associated protein 1 lightchain LC3 — LC3II. It was also accompanied by the inhibition of heat shock proteins Hsp27 and Hsp72 expression.Our results indicate that imperatorin alone, or in combination with cisplatin, is mainly an autopahgy inducerin HeLa cells.There is growing evidence that commonly applied chemotherapy regimens can be improved by introducingnew, specific, active and low side-effect drugs, or by combining substances to obtain the required clinicaleffect. The aim of the present study was to investigate the effects of imperatorin and cisplatin, applied separatelyor in combination, on apoptosis, necrosis and autophagy induction in the human cervical carcinoma cell line(HeLa). Imperatorin appeared to be a potent autophagy inducer, rather than a necrotic or apoptotic one. Incontrast, cisplatin induced mainly apoptosis and necrosis after 6 h and 24 h, while longer incubation resultedonly in necrosis induction. When HeLa cells were incubated with both drugs, autophagy appeared most frequently,although to a smaller extent than that observed after imperatorin administered alone. At the molecularlevel, autophagy was correlated with the presence of the cleaved form of microtubule-associated protein 1 lightchain LC3 — LC3II. It was also accompanied by the inhibition of heat shock proteins Hsp27 and Hsp72 expression.Our results indicate that imperatorin alone, or in combination with cisplatin, is mainly an autopahgy inducerin HeLa cells

The role of collagen in co-cultures of human normal corneal and conjunctival cells

Background: Intermediate interactions between corneal and conjunctival epithelial cells play an important rolein the process of correct vision. The goal of this paper was to establish whether the presence or absence of collagentype I changes paracrine interactions between corneal and conjunctival epithelial cells. Material and methods: Cultures of human corneal and conjunctival epithelial cells were used in the study. TheELISA quantitative analysis of interleukin 1β (IL-1β), interleukin 6 (IL-6), urokinase-type plasminogen activator(uPA), and uPA receptor (uPAR), assessment of the type of interactions between cells, as well as correlations betweentested parameters were performed. Results: The presence of collagen type I changed the quantitative production of IL-1β and IL-6 by the examinedcells in the co-culture system. It did not affect the level of released uPA and uPAR. The presence or absence ofcollagen also changed the relationship between the cells, which were evaluated in relation to changes in the level ofreleased cytokines. Conclusions: Different levels of collagen type I constituting a component of extracellular matrix proteins significantlyaffect and regulate the indirect interactions between human corneal and conjunctival epithelial cells

Application of primary cell cultures of laryngeal carcinoma and laser scanning cytometry in the evaluation of tumor reactivity to cisplatinum.

Unsatisfactory effects of treatment of laryngeal carcinoma patients stimulate the clinicians as well as researchers to develop new more effective treatment models and to find new reliable prognostic factors. The aim of the present study was the evaluation of the use of primary cell cultures of the laryngeal carcinoma and laser scanning cytometry (LSC) in the assessment of tumor reactivity to cisplatinum. Nineteen primary cultures of laryngeal carcinoma cells established from fragments of laryngeal carcinoma infiltrations were cultured with or without cisplatin, stained with monoclonal antibodies against P53 and BCL-2 proteins and analyzed by LSC. Cisplatin added to the culture medium leads to the significant increase of P53 expression and decrease of BCL-2 expression. Moreover, changes of P53 and BCL-2 expressions were significantly correlated. Our findings of apoptosis regulatory mechanisms could be useful in patient qualification for the chemotherapeutic follow-up treatment

Effect of diosmin and diosmetin on the level of pro-inflammatory factors in the endothelium artificially induced with inflammatory stimuli

Introduction: Diosmin and its aglycone diosmetin are phlebotropic drugs used in the treatment of chronic venous insufficiency (CVI). Diosmin increases the elasticity and tension of blood vessel walls, exhibits an antiedematous effect, and acts as an anti-inflammatory agent. As it is commonly known that the endothelium layer plays a significant role in the physiology and pathophysiology of the cardiovascular system, this paper investigates the effect of diosmin and diosmetin on modulating the levels of pro-inflammatory factors in an endothelial cell culture (HUVEC) stimulated by lipopolysaccharide (LPS) or phorbol (PMA). Material and methods: A normal human umbilical vein/vascular endothelium cell line HUV-EC-C (HUVEC) was stimulated with lipopolysaccharide (LPS) or phorbol 12-myristate-13-acetate (PMA). Cell viability was assessed using NR and MTT assays. The levels of human IL-1β, IL-6, IL-10, COX-2, and PGE2 were measured using ELISA kits. Results: Depending on the agent used to initiate inflammation, different levels of factors associated with this state were obtained. Diosmetin significantly decreased the levels of pro-inflammatory IL-1β and IL-6 as well as COX-2 in PMA-treated cells. Meanwhile, diosmin did not affect the interleukins but it lowered COX-2 and increased PGE-2. Upon the LPS stimulation of HUVEC cells, diosmetin increased the levels of PGE2, IL-1β, COX-2, and nitric oxide (NO), while diosmin increased NO and IL-6. Conclusion: Diosmin and diosmetin have different impacts on the levels of pro-inflammatory factors depending on the inflammation inducer. Diosmetin more effectively modulated inflammation than diosmin, suggesting that the attachment of the sugar moiety to the aglycone attenuates its activity

Homocysteine influences the human keratocytes cell cycle

BACKGROUND: Homocysteine (Hcy), a metabolic intermediate, is a sulfur-containing amino acid not present in the structure of proteins. It has been shown that high Hcy levels via oxidative stress, induced inflammation, and vessel dysfunction may affect the functioning of tissues, including the structures that form the eye, among others keratocytes. The visual disturbance caused by high levels of Hcy may also be associated with disturbed cell proliferation resulting from the effect of this amino acid on the cell cycle. The goal was to analyse the influence of Hcy on the keratocytes and to find out in which phase of the cell cycle its course is disturbed by this amino acid. MATERIALS AND METHODS: A normal human keratocytes (HK) cell line was used in the study. May-Grünwald-Giemsa (MGG) staining for morphology visualization and cytometric cell cycle analysis were performed. RESULTS: Hcy does not affect the G1 phase of the cycle, while it regulates the S and G2 phases. Changes in the amount of the sub-G1 population  indicative of a pro-apoptotic effect of Hcy on keratocytes were detected. The form of the Hcy administered (L stereoisomer or DL racemic mixture) and the amino acid concentration were also important. CONCLUSIONS: Homocysteine influences the keratocyte cell cycle. The change occurs at the stage of the G1-S transition, which suggests a decreasing level of cells in the S phase and an increasing level in the G2 phase. The long-term influence of Hcy on keratocytes may affect keratocytes proliferation and possible cornea regeneration

In Vitro Antiproliferative and Antioxidant Effects of Extracts from Rubus caesius

The present study was performed to evaluate the effect of different extracts and subfractions from Rubus caesius leaves on two human colon cancer cell lines obtained from two stages of the disease progression lines HT29 and SW948. Tested samples inhibited the viability of cells, both HT29 and SW948 lines, in a concentration-dependent manner. The most active was the ethyl acetate fraction which, applied at the highest concentration (250 μg/mL), decreased the viability of cells (HT29 and SW948) below 66%. The extracts and subfractions were also investigated for antioxidant activities on DPPH and FRAP assays. All extracts, with the exception of water extract at a dose of 250 μg/mL, almost totally reduced DPPH. The highest Fe3+ ion reduction was shown for the diethyl and ethyl acetate fractions. It was more than 6.5 times higher (at a dose 250 μg/mL) as compared to the control. The LC-MS studies of the analysed preparations showed that all samples contain a wide variety of polyphenolics, among which ellagitannins turned out to be the main constituents with dominant ellagic acid, sanguiin H-6, and flavonol derivatives