Protective effects of Thymoquinone against Methotrexate-Induced germ cell apoptosis in male mice

Background: Toxic effects of anti-cancer and other drugs on the normal tissues could be reduced by the herbal plants and their fractions. This study investigated the protective effect of thymoquinone [TQ] as a fraction of Nigella sativa on methotrexate [MTX]- induced germ cell apoptosis in male mice

Materials and Methods: In this experimental study, thirty male Balb/c mice were divided randomly into 5 groups [n=6]. A single dose of MTX [20 mg/kg] and different concentrations of TQ were administrated for 4 consecutive days. Terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL] assay was performed on paraffin embedded tissue sections to analysis the occurrence of apoptosis in the testis. Reverse transcription polymerase chain reaction [RT-PCR] of apoptosis-related genes was performed with RNA extracted from testes of the mice. Statistical analysis was done using one-way ANOVA

Results: In the MTX group, there was a significant increase in morphologic sign of germ cell degeneration of tubules [48 +/- 0.6%], apoptotic index [AI; 2.3 +/- 0.6%], as well as mRNA expression of p53 [P=0.008], caspase 8 [P=0.002], caspase 3 [P=0.005], caspase 9 [P=0.000], bax [P=0.004] and the ratio of bax/bcl-2 [P=0.000], whereas there was an decrease in the expression of bcl-2 [P=0.003], as compared to control group. In MTX+TQ groups, the data showed that different concentrations of TQ could improve the harmful effects caused by the MTX. The best protective effects were achieved in MTX+TQ [10 mg/kg]

Conclusion: TQ protects testicular germ cell against MTX-induced apoptosis by affecting related genes regulation

Effects of Tamoxifen in Combination with Tranilast on CXCL12-CXCR4 Axis and Invasion in Breast Cancer Cell Lines

It has been reported that CXCL12 binding to CXCR4 induces several intracellular signaling pathways, and enhances survival, proliferation, and migration of malignant cells. In the present study, we examined the effects of anti-estrogen tamoxifen and anti-allergic tranilast drugs as a single or in combination on invasion by two in-vitro invasion assays. wound-healing and matrigel invasion on MCF-7 and MDA-MB-231 human breast cancer cell lines. The mRNA expression levels of CXCR4 and CXCL 12 were measured by quantitative real time-RT PCR and CXCL 12 protein levels were evaluated by ELISA assay. The data showed that treatment with tamoxifen and tranilast as a single or in combination resulted in decreased CXCR4 and CXCL 12 mRNA and CXCL 12 protein expression levels. Both in-vitro invasion assays markedly showed synergistic effect of tamoxifen when combined with tranilast drug. Either ER-positive or ER-negative breast cancer cells were sensitive to this combination therapy. In conclusion, Tranilast increases antimetastatic effect of tamoxifen. The synergistic effect of tranilast is not estrogen dependent; however tamoxifen may sensitize the cells for the action of tranilast. The data also support the importance of the CXCR4/CXCL12 interaction in breast cancer metastasis, and further suggest that CXCR4 and CXCL 12 are critical targets for tamoxifen and tranilast in combination or alone

Synthesis, and in-vitro cytotoxicity studies of a series of triazene derivatives on human cancer cell lines

Compounds containing triazene ring structure are cytotoxic agents and clinically used as antitumor alkylating agents. In this study, a series of triazene derivatives holding alkyl and aryl moieties were synthesized and proved to be potent cytotoxic agents in-vitro particularly against eight cancer cell lines [PCS, HT29, Hela, HL60, Jurkat, K562, MCF7, HepG2] and a non-cancerous cell line [HUVEC]. The cytotoxic activity was assessed using two methods, LDH assay, and trypan blue exclusion. Some of the triazene derivatives showed cytotoxic activity more than temozolomide [TMZ] as the reference drug. The synthesized triazenes showed marked cytotoxicity effects on all eight cancer cell lines. Among the compounds synthesized, l,3-bis [2-ethoxyphenyl] triazene C had unique efficacy and selectivity so that it had IC[50] between 0.560-3.33 microM on cancer cell lines and 12.61 microM on normal cell line [HUVEC]. l-[4-nitrophenyl]-3-[2-hydroxyethyl] triazene E shows weaker effect on cancer cell lines than the other compounds having 1C[50] between 3-15.54 microM

[Primary culture of human skin melanocyte and comparison of culture in the presence and absence of phorbol ester]

Primary culture takes place following the cell isolation from tissues. Isolation and culture of melanocytes based on their roll in the protection of body against hazardous sun rays, production of skin, cornea and hair color is really important. This study was done to set isolation, culture and proliferation of melanocytes from children foreskin and adult eyelashes, and also comparison of two types of melanocyte culture medium. Human foreskin and eyelash samples were used for melanocyte isolation and culture. After isolation of epidermis from dermis, epidermis cell suspensions were prepared by enzymatic digestion. The isolated cells were cultured in two melanocyte selective culture media. Immunocytochemistary and reverse transcription-polymerase chain reaction [RT-PCR] assays were used for confirmation of isolated and cultured melanocytes. Our results indicated that isolated melanocyte cultured in the selective medium without phorbol esters is better than the melanocytes cultured in selective medium containing phorbol esters not only morphologically but also physiologically and from the aspect of cell adhesion. In addition, the results showed that isolated melanocyte from adult eyelashes are more dendritic than melanocytes isolated from children foreskin. Conversely, our results indicated that the number of cell passages in melanocyte isolated from foreskin is more than melanocytes isolated from adult eyelashes. Melanocytes cultured in selective medium containing convenient growth factors in absence of phorbol esters show more native physiological and adhesive properties. In addition, melanocyte isolated from younger tissues such as foreskin have better proliferative and sub-culturing properties so we suggest isolation and culture of younger tissues.

Study of the mechanisms and causes related to angiogenesis

Angiogenesis, the process of new blood vessels formation from existing ones, is important for the normal body growth and development. Angiogenesis is dependent on a delicate equilibrium between endogenous angiogenic and anti-angiogenic factors. If this equilibrium is disturbed, different diseases such as corneal neovascularization, endometriosis, obesity, atherosclerosis, diabetic retinopathy, psoriasis, tumor growth and metastasis may ensue. In general, angiogenesis process is under the influence of several factors and consists of cellular events such as migration, proliferation and differentiation of endothelial cells and finally formation, maturation and remodeling of vessels. Hence, angiogenesis inhibition as a useful measure in conventional therapies such as chemotherapy and radiation has attracted scientists' attention studying on these issues. In recent years, the angiogenesis control has been considered as a novel idea for control and treatment of angiogenesis-dependent disorders especially tumor growth and metastasis. Because of its importance in causing these disorders, we discuss various aspects of angiogenesis, its mechanisms and causes and also the related studies in this review article

Anti tumor activity of salvia officinalis is due to its anti-angiogenic, anti-migratory and anti-proliferative effects

The significant factor contributing to the distant invasion of cancer cells is the ability of tumors to produce large numbers of new blood vessels, known as angiogenesis. Many natural products inhibit angiogenesis. Herein, ethanol extract of Salvia officinalis [SO] has been analyzed for its anti-angiogenic, anti-proliferation and anti-migration activities. The anti-angiogenic effect of the SO extract was evaluated on chicken chorioallantoic membrane [CAM] neovascularisation model, microscopically. The inhibitory effect of the extract on human umbilical vein endothelial cells [HUVECs] migration was tested on the wound-healing model with an inverted microscope. In addition, SO extract was screened for its possible anti-proliferative effects by separately counting HUVECs, Wehi and K562 cells with cell counter against their control wells, So extract exhibited a significant inhibitory activity in CAM assay in a dose dependent manner. CAM angiogenesis was gradually prevented to from at 100 micro g/ml of SO extract, but completely inhibited to form at 200 micro g/ml. After human umbilical vein endothelial cells [HUVECs] were suppressed by dose-dependent SO extract, their migrations were detected by wound healing model, yet they were unable to show a dose response effect on proliferation of the different cells [50-200 micro g/ml]. As observing in this study, SOextract could inhibit proliferation of the different cells at the concentrations above 200 micro g/ml without toxic effect on the cells in doses ranged from 0-500 micro g/ml. These findings indicated that SO extract might be a promising candidate for anti-angiogenic treatment

Anti-angiogenic activity of ficus carica latex extract on human umbilical vein endothelial cells

Angiogenesis, the formation of new blood vessels, is a key process in cancer development and metastasis. In this study, the anti-angiogenic and anti-proiiferative potentials of Ficus carica latex extract have been investigated using human umbilical vein endothelial cells [HUVECs]. Different doses of latex extract were prepared and added to a three-dimensional culture of HUVEC in a collagen matrix. After 3-5 days of treatment, the anti-angiogenic effects of the extracts were monitored microscopically. For the anti-proliferation assay, different doses of the extracts were examined on HUVECs. The results clearly indicated that latex extract could inhibit proliferation and capillary tube formation of HUVECs in a dose-dependent manner at the range of 100-400 micro g/ml. In addition, the extract was not cytotoxic up to 450 micro g/ml as assessed by trypan blue and lactate dehydrogenase [LDH] cytotoxicity assays. It is concluded that latex extracts of F. carica contain strong anti-angiogenic and anti-proliferative activities. Our data indicates that latex extract could be a candidate as a potential agent for the prevention of angiogenesis in cancer and other chronic disorders

[Angiogenesis and tumor]

Angiogenesis, the process of new blood vessel formation from existing ones, plays an important role in the physiologic circumstances such as embryonic development, placenta formation, and wound healing. It is also crucial to progress of pathogenic processes of a variety of disorders, including tumor growth and metastasis. In general, angiogenesis process is a multi-factorial and highly structured sequence of cellular events comprising migration, proliferation and differentiation of endothelial cells and finally vascular formation, maturation and remodeling.Thereby, angiogenesis inhibition as a helping agent to conventional therapies such as chemotherapy and radiation has attracted the scientists' attentions studying in this field

Human coagulated plasma as a natural and low cost matrix for in vitro angiogenesis

Angiogenesis, the development of new blood vessels, is an important process in tissue development and wound healing, but becomes pathologic when associated with solid tumor growth, proliferative retinopathies, and rheumatoid arthritis. Accurate and reliable qualification of neovascular [angiogenic] response, both in vitro and in vivo, is an essential requirement for the study of new blood vessel growth. The complexity of currently used three-dimensional in vitro angiogenesis systems makes it difficult to approve material in its models. Capillary-like structure occurs on basement membrane components such as collagen and/or laminin, while in other models, CLS formation occurs on transitional matrices such as fibrin. To solve this problem, we were interested in developing an angiogenesis system which allows rapid and reliable quantification of three-dimensional neovessel formation in vitro. Human bone marrow endothelial cells were seeded on gelatin-coated microcarriers and suspended in a solution of platelet-poor plasma which was induced to polymerize by addition of calcium chloride. In this way, microcarriers were entrapped in three-dimensional coagulated plasma. Within a few hours, endothelial cells begin to leave these supporting microcarries and migrate into the coagulated-plasma matrix and formed CLS within 48-72 hours. We developed a convenient angiogenesis in vitro system which allows reliable quantification of capillary formation in a three-dimensional environment