Anticancer potential of bovine lactoferrin in colorectal and other cancers - a brief overview

Colorectal cancer (CRC) is the second leading cause of cancer-related death in the world, thus constitutes a serious medical challenge. Due to the high mortality and also large number of recurrences of the disease, great emphasis is placed on the screening of people from high risk groups as well as the use of chemopreventive agents in chronic inflammation of the intestine. Chemoprevention is an early intervention decreasing risk of cancer development. Term chemoprevention comes from 1976 and is explained as “use of a synthetic or natural substance to decrease the risk of cancer developing, delay the time of cancer onset, or to reverse the carcinogenesis process.” Lactoferrin (Lf) is a glycoprotein, which is included as a member of the transferrin family. It is able to reversibly bind iron ions, thus participating in maintaining iron homeostasis in the body. Lf mainly appears in the milk of most mammals, but also occurs in different biological fluids like tears, saliva, mucus and bronchial secretions. Lactoferrin is a multipotent molecule which exerts animicrobial, antiviral, antiparasitic and antifungal activity. Moreover, Lf has anti-inflammatory, immune regulatory and anticancer properties. Lf increases the immune system ability to eliminate cancer cells, thus preventing cancer development. Lf treatment was also found efficient in inhibiting proliferation, migration, survival and induction programming cell death in cancer cells. The present paper aims to review the molecular basis of lactoferrin chemopreventive effect in colorectal cancer. Bovine lactoferrin (bLf) acts both on stimulating immunological anticancer response and also directly through inducing cell cycle arrest, apoptosis or inhibition cell migrations. Moreover Lf boosts the effect of chemotherapy and alleviates its side effects. Finally, Lf is a nutraceutical characterized by high selectivity toward cancer cells and simultaneously has a high safety profile. It has considerable potential in the fight against CRC in both prevention and treatment

Substances with therapeutic potential affecting the Keap1-Nrf2-ARE pathway in age-related eye diseases

One of the features of the uninterrupted progress of civilization, also in the field of medicine, is the extension of the average life length, which ultimately leads to the aging of societies in developed and developing countries. The disadvantage of this phenomenon is the higher appearance of age-related diseases. These include, among others, eye diseases such as: cataracts, glaucoma, diabetic retinopathy or age-related macular degeneration, which are the most common reasons for the irreversible loss of vision among elderly people. However, age is not the only risk factor for developing these diseases. Among others, we can distinguish: lifestyle, use of stimulants such as alcohol, nicotine, exposure to UV radiation or accompanying diseases such as diabetes. However, a common factor in their development is oxidative stress, and the eye is an organ particularly sensitive to its effects. Especially the retina and the cells of the retinal pigment epithelium are exposed to an extremely high load of oxidative stress, which increases with age. The human body has many antioxidant defences mechanisms, and one of them that plays an important role ,also in the antioxidant mechanisms of the eye, is the the Keap1-Nrf2-ARE signalling pathway. Nrf2 is a nuclear transcription factor of erythroid origin, type 2, which is present in the cytosol in the form of a complex with its inhibitor, the Keap1 protein. Under normal, non-stress conditions, Nrf2 is destined for proteasomal degradation, but under oxidative stress it is dissociated and transduced into the cell nucleus. As a result, Nrf2 activates the expression of genes known as elements of the antioxidant response. They are responsible for the synthesis of phase II cytoprotective enzymes, which include: heme oxygenase 1, superoxide dismutase, glutathione S-transferase or γ-glutamyl cysteine ligase. Due to the fact that oxidative stress is involved in the development of many diseases, signalling pathways that reduce its level may be a potential therapeutic target. Especially the multifaceted influence on the functioning of the Nrf2 factor as ubiquitous in the body and modifications of the Keap1-Nrf2-ARE signalling pathway seem to be a promising direction in the search for new forms of therapy, also in the case of age-related eye diseases. Among the numerous substances with therapeutic potential that affect the activity of the Keap1-Nrf2-ARE pathway, plant-derived compounds such as lutein, zeaxanthin, polyphenols such as quercetin and resveratrol, and drugs such as probucol and calcium dobesylate should be mentioned

Transformujący czynnik wzrostu beta w raku podstawnokomórkowym, kolczystokomórkowym i rogowiaku kolczystokomórkowym

Introduction. Transforming Growth Factor β (TGFβ) activates signaling cascades which regulate cell proliferation, differentiation, apoptosis, inflammatory response and angiogenesis. In the early stages of malignant transformation this cytokine acts as an inhibitor of tumour growth. In the advanced stages of malignant transformation TGFβ acts as a promoter of metastasis. Changes in the expression of genes associated with TGFβactivity could provide a new strategy of molecularly targeted therapy.Aim. The aim of this study was to compare the mRNA profile of genes associated with TGFβ signaling pathways in non-melanoma skin pathologies biopsy specimens of bas-al cell carcinoma (BCC), squamous cell carcinoma (SCC) and keratoacanthoma (KA) in comparison to normal skin.Material and methods. Tissue samples of KA, SCC and BCC were obtained from the central part of tumours. Healthy skin margins comprised the control group. mRNA profile of genes coding TGFβ and proteins involved in TGFβ-induced signaling pathways was determined using oligonucleotide microarrays (Affymetrix).Results. Microarray analysis showed changes in profile of genes coding proteins in-volved in TGFβ-induced signaling pathways. In SCC TGFβ-1 (TGFB1) was upregulated, comparing to controls. Both in KA and SCC, the most statistically significant change re-ferred to TGFBR3 (Transforming Growth Factor beta Receptor III) mRNA.Conclusions. mRNA profile of genes coding proteins involved in TGFβ-induced signal-ization reveals strong molecular similarity of SCC and KA

Poziom ekspresji defensyny DEFB4A w różnicowaniu rogowiaka kolczystokomórkowego, raka kolczystokomórkowego i raka podstawnokomórkowego

Introduction. Defensins are peptide with antimicrobial, antiviral, antifungal activities and many other functions, such as induction of immunological response and antitumor activity. Changes in expression level of defensins was studied in many skin pathologies, including dermatological lesions such as psoriasis, atopic dermatitis and non-melanoma skin cancers (squamous cell carcinoma – SCC and basal cell carcinoma – BCC).Aim. The objective of this study was to evaluate the mRNA profile of defensin-related genes’ transcripts as an additional molecular marker of non-melonoma skin pathologies: SCC, BCC and keratoacanthoma (KA).Material and methods. Tissue samples were obtained from the central part of tumours (KA, SCC and BCC) and healthy margins. mRNA profile of genes coding defensins and proteins involved in their activation was determined using oligonucleotide microarrays (Affymetrix). Vali-dation of the microarray analysis was performed using real-time QRT-PCR.Results. Microarray analysis revealed changes in defensin-related genes’ profile. In all tumours DEFB4A (defensin beta 2) mRNA was up-regulated, compared with the healthy skin margins. Real-time QRT-PCR analysis showed increased DEFB4A transcript level both in KA and SCC comparing to BCC.Conclusions. Defensin beta 2 mRNA level is a useful tool for the differentiation of KA and SCC from BCC. KA and SCC cannot be differentiated on the basis of the DEFB4AmRNA level

Pharmacological and non-pharmacological methods of psoriasis therapy with particular emphasis on biopharmaceuticals

Psoriasis is a chronic, inflammatory and recurrent skin disease of an autoimmune nature. Genetic, behavioral and environmental factors influence the onset, course and severity of skin lesions. Precursor factors conditioning the formation of psoriatic lesions may include, among others, strong psychological stress, trauma, acute infection - mainly infections caused by Streptococcus species, surgery, hormonal changes, as well as alcohol abuse, smoking, and taking certain medications. In the course of psoriasis, increased proliferation of mainly keratinocytes is observed at the tissue level, while increased secretion of proinflammatory cytokines and adipokines is observed molecularly. The goal of psoriasis treatment is to reduce clinical changes, prolong remission, prevent complications, and restore the patient's life and professional fitness. The choice of treatment should enable control of the course of the disease while reducing side effects that may negatively affect the patient's life. Due to, among others, the genetic conditions underlying the disease, complete recovery is not possible, resulting in frequent relapses. The choice of method depends on the severity of skin lesions, the frequency of relapses and the course of the disease. In order to assess the severity of psoriatic lesions and choose the appropriate treatment, measuring scales are used, which are mainly based on the physician's visual assessment of psoriatic lesions and the patient's subjective feelings. However, one correct treatment regimen cannot be demonstrated, and each therapy should be individually tailored to the patient. In the treatment of psoriasis, PUVA photo chemotherapy, UVB irradiation, photo protection, glucocorticosteroids, retinoids, vitamin D analogues, reducing agents, keratotic agents, and immunosuppressant - cyclosporin, methotrexate and hydroxycarbamide as well as biological therapy affecting specific mediators of immune reactions are used. Unfortunately, despite the molecularly targeted treatment, the phenomenon of drug resistance is observed.Łuszczyca jest przewlekłą, zapalną i nawracającą chorobą skóry o charakterze autoimmunologicznym. Na początek, przebieg i nasilenie zmian skórnych wpływają czynniki genetyczne, behawioralne i środowiskowe. Czynnikami prekursorowymi warunkującymi powstawanie zmian łuszczycowych mogą być między innymi: silny stres psychiczny, uraz, ostra infekcja - głównie zakażenia wywołane przez Streptococcus species, zabieg operacyjny, zmiany hormonalne, a także nadużywanie alkoholu, palenie papierosów, czy zażywanie niektórych leków. W przebiegu łuszczycy obserwuje się na poziomie tkankowym nasiloną proliferację głównie keratynocytów, natomiast molekularnie stwierdza się wzmożoną sekrecję cytokin prozapalnych i adipokin. Celem leczenia łuszczycy jest redukcja zmian klinicznych, wydłużenie remisji, zapobieganie powikłaniom oraz przywrócenie sprawności życiowej i zawodowej chorego. Wybór terapii powinien umożliwiać kontrolę przebiegu choroby, jednocześnie redukując skutki uboczne mogące negatywnie wpływać na życie chorego. Ze względu między innymi na uwarunkowania genetyczne będące podłożem choroby nie jest możliwe całkowite wyleczenie, co skutkuje częstymi nawrotami. W celu oceny stopnia nasilenia zmian łuszczycowych oraz wyboru odpowiedniej terapii stosuje się skale pomiarowe, opierające się głównie na wizualnej ocenie zmian łuszczycowych przez lekarza oraz subiektywnych odczuciach pacjenta. Nie można jednak wykazać jednego słusznego schematu leczenia, a każda terapia powinna być indywidualnie dobierana do chorego. W leczeniu łuszczycy stosuje się: fotochemioterapię PUVA, naświetlanie UVB, fotoprotekcję, glukokortykosteroidy, retinoidy, analogi witaminy D, środki redukujące, środki keratolityczne, środki immunosupresyjne- cyklosporynę, metotreksat i hydroksykarbamid oraz terapię biologiczną oddziałującą na specyficzne mediatory reakcji immunologicznych/zapalnych. Niestety coraz częściej pomimo molekularnie ukierunkowanego leczenia obserwuje się zjawisko lekooporności

Zmiany w ekspresji genów kodujących białka związane z aktywnością kaspaz oraz białka z rodziny BCL-2 w komórkach RPTEC traktowanych amfoterycyną B i jej modyfikowanymi formami

INTRODUCTION: The main limitation of the use of amphotericin B (AmB) – effective in the treatment of systemic fungal infections – is its high toxicity to human cells. The mechanism of AmB toxicity is not clear. Caspase-related and BCL-2 proteins participate in the regulation of apoptosis. Thus, they may be involved in drug toxicity. In this study we evaluated the influence of AmB on the transcriptional activity of genes related to caspases and the BCL-2 family. We also tested the influence of modified forms of AmB: AmB-Cu2+ (the complex with copper(II) ions) and the AmB-ox (oxidized form). MATERIAL AND METHODS: Human RPTECs (Renal Proximal Tubule Epithelial Cells) were treated with AmB, AmB-Cu2+ and AmB-ox. Total RNA was extracted using the phenol-chloroform method. The expression profiles of genes related to caspase activity and BCL-2 were determined using oligonucleotide microarrays (HG-U133A 2.0, Affymetrix). Analysis included 67 ID related to caspases and 32 ID associated with BCL-2, according to the Affymetrix database. RESULTS: The analysis revealed upregulation of the BCL-2 and BCL2L1genes in the cells treated with AmB-Cu2+, in comparison to the control. In both the AmB and AmB-Cu2+ -treated cells, differentiating genes were associated with inflammation and mitophagy activated by intrinsic signals. In the cells treated with AmB-ox, the BCL-2 genes were downregulated. CONCLUSIONS: The results suggest that AmB and AmB-Cu2+ activate genes involved in the regulation of inflam-mation and autophagy induced by intrinsic signals, but overexpression of BCL-2 and BCL2L1 may protect AmB-Cu2+--treated cells from death. In the cells treated with AmB-ox extrinsic signals prevail, indicating the distinct molecular mechanism of its cytotoxicity.WSTĘP: Głównym ograniczeniem stosowania amfoterycyny B (AmB) – skutecznej w leczeniu grzybic układowych – jest jej wysoka toksyczność wobec komórek ludzkich. Mechanizm cytotoksyczności nie został wyjaśniony. Białka związane z aktywnością kaspaz oraz białka należące do rodziny BCL-2 uczestniczą w regulacji apoptozy, mogą być zatem zaangażowane w procesy odpowiedzialne za toksyczność leku. W pracy oceniono wpływ AmB na aktywność transkrypcyjną genów kodujących białka związane z aktywnością kaspaz oraz białka z rodziny BCL-2. Zbadano również wpływ modyfikowanych form AmB: AmB-Cu2+ (kompleks z jonami miedzi (II)) i AmB-ox (formy utlenione). MATERIAŁ I METODY: Ludzkie komórki RPTECs (Human Renal Proximal Tubule Epithelial Cells) inkubowano z AmB, AmB-Cu2+ i AmB-ox. Całkowity RNA wyekstrahowano metodą fenolowo-chloroformową. Profil ekspresji genów wyznaczono techniką mikromacierzy oligonukleotydowych (HG-U133A 2.0, Affymetrix). Analiza obejmowała 67 ID genów związanych z aktywnością kaspaz i 32 ID geny kodujące białka z rodziny BCL-2, zaproponowane przez bazę Affymetrix. WYNIKI: Analiza wykazała nadekspresję genów BCL-2 i BCL2L1 w komórkach traktowanych AmB-Cu2+, w porównaniu z kontrolą. Zarówno w komórkach traktowanych AmB, jak i AmB-Cu2+ geny różnicujące związane były z za-paleniem i mitofagią aktywowanymi w odpowiedzi na sygnały wewnątrzkomórkowe. W komórkach traktowanych AmB-ox geny z rodziny BCL-2 były wyciszone. WNIOSKI: Wyniki sugerują, że AmB i AmB-Cu2+ aktywują geny zaangażowane w regulację zapalenia i mitofagii aktywowanych sygnałami wewnątrzkomórkowymi, jednak nadekspresja genów BCL-2 i BCL2L1 może chronić komórki traktowane AmB-Cu2+ przed śmiercią. W komórkach traktowanych AmB-ox przeważa sygnał zewnątrzkomórkowy, co wskazuje na odrębny mechanizm cytotoksyczności tej formy antybiotyku

Influence of elastin-derived peptides, glucose, LDL and oxLDL on nitric oxide synthase expression in human umbilical artery endothelial cells

Endothelial dysfunction plays an important role in the development of atherosclerosis. Elastin-derived peptides (EDP), hyperglycemia, hypercholesterolemia and oxidized LDL have a proven proatherosclerotic potential. Nitric oxide generated by endothelial nitric oxide synthase (eNOS; EC 1.14.13.39) is an important vasorelaxant. Here we studied the influence of those proatherosclerotic factors on eNOS gene and protein expression in artery-derived endothelial cells. Human umbilical artery endothelial cells (HUAEC) were incubated with or without: glucose (270 mg/dl), LDL (200 mg/dl), oxidized LDL (oxLDL 25 mg/dl) or κ-elastin (0.5 and 2.5 µg/ml). Gene expression was assessed by real time RT-PCR, whilst the eNOS protein by ELISA. In cells incubated with 2.5 µg/ml of κ-elastin, a 31 % increase of eNOS mRNA expression was observed, but the protein level remained unchanged. OxLDL, LDL and glucose decreased the eNOS protein level by 74 %, 37 % and 29 %, respectively. OxLDL decreased eNOS mRNA by 42 %. LDL non-significantly decreased eNOS mRNA expression. An elevated glucose level did not affect the eNOS mRNA expression. Hyperglycemia and an elevated level of LDL, particularly oxLDL, decreased the level of eNOS protein in endothelial cells. As κ-elastin did not decrease the expression of eNOS gene in HUAEC, the proatherogenic properties of elastin-derived peptides are unlikely to be due to their influence on eNOS