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Background and Aims: Previous studies have shown that adipose-derived mesenchymal stem/ stromal cells are one of the sources of mesenchymal stem cells (MSCs) with the capacity to differentiate into various mesodermal cell lineages. MSCs with cytokines secretion capability, which contributes to repair damaged tissues have gained wide credence for future cell-based therapeutic applications. In this study, the effect of the different dosages of vitamin E and Selenium was assessed on the stemness of the human adipose tissue-derived MSCs (AD-MSCs). Materials and Methods: Following 24 hours of cell treatments with different dosages of vitamin E and Selenium, MTT assay was used to assess the effect of them on cell proliferation. Moreover, the stemness of the AD-MSCs was assessed using osteogenic and adipogenic induction medium supplemented with the different dosages of the vitamin E and Selenium. Finally, Alizarin red and Oil-red O staining were performed to detect matrix mineralization and lipid droplet accumulation, respectively. Results: MTT data revealed that the optimal concentration for vitamin E and Selenium were 125 µM and 121 µM for the viability of the AD-MSCs. Moreover, the effect of vitamin E and Selenium were assessed by osteogenic and adipogenic differentiation by optimal dosages obtained by MTT assay, respectively. Maximum mineralization and lipid droplet aggregation of the differentiated cells were detected at IC50 in comparison with the control group. Conclusions: These results suggest that different dosages of vitamin E and Selenium could have various impacts on the proliferation and differentiation induction of human AD-MSCs.

The low solubility of the plant-extracted agent like D-limonene in cancer therapy is a critical problem. In this study, we prepared D-limonene-loaded niosomes (D-limonene/Nio) for cancer therapy through in vitro cytotoxicity assay of HepG2, MCF-7, and A549 cell lines. The niosomal formulation was prepared by film hydration technique with Span® 40: Tween® 40: cholesterol (35:35:30 molar ratio) and characterized for vesicle distribution size, morphology, entrapment efficiency (EE%), and in vitro release behaviour. The obtained niosomes showed a nanometric size and spherical morphology with EE% about 87 ± 1.8%. Remarkably prolonged release of D-limonene from niosomes compared to free D-limonene observed. The loaded formulation showed significantly enhanced cytotoxic activity with all three cancer cell lines (HepG2, Macf-7 and A549) at the concentration of 20 μM. These results indicated that niosome loaded with phytochemicals can be a promising nano-carrier for cancer therapy applications

A variety of biological activities, such as anti-microbial and anti-tumor properties was reported for 1,10-phenanthroline and its copper complexes. In this study, the anti-proliferative activity of a novel [Cu(L)(phen)] complex was investigated on MCF-7 breast cancer cells using MTT assay. Since chemotherapy is lake of ability to distinguish between normal cells from cancerous cells, therefore we also investigated the effect of [Cu(L)(phen)] complex on normal L929 cells. The results showed that following 24 and 48 h exposure of cells with [Cu(L)(phen)] complex, the IC50 values for MCF-7 were significantly lower than that recorded for L929 and normal cells were less sensitive than cancerous cells to the complex. Additionally, the [Cu(L)(phen)] complex displayed a time- and concentration-dependent cytotoxic response, with MCF-7 and L929 cells. Also flow cytometry findings suggest that [Cu(L)(phen)] complex is capable of decreasing cancer cell viability through apoptosis and did not efficiently activate the necrosis process.

BACKGROUND: The use of phytochemicals to prevent or suppress tumours is known as chemoprevention. Numerous plant-derived agents have been reported to have anticancer potentials. As one such anticancer phytochemical, diosgenin has several applications which are nevertheless limited due to its low solubility in water. METHODS: We loaded diosgenin into niosome to increase its solubility and hence efficiency. Diosgenin-niosome (diosgenin loaded into niosome) was prepared by thin-film hydration method and characterised by optical microscopy, dynamic light scattering (DLS), scanning electron microscopy (SEM), and UV-visible spectrophotometry. Also, loading efficiency, in vitro drug release, and cytotoxicity assay were performed on HepG2 cell line. RESULTS AND DISCUSSION: Diosgenin-niosome has a nanometric size with a normal size distribution and spherical morphology. The loading efficiency of diosgenin was about 89% with a sustainable and controllable release rate. Finally, the viability of free diosgenin was 61.25%, and after loading into niosomes, it was improved to 28.32%. CONCLUSION: The results demonstrated that niosomes increase the solubility of naturally derived hydrophobic chemicals and thus enhance their anticancer effect. [Figure: see text]

Ubiquitination is an important cellular mechanism with a pivotal role in the degradation of abnormal or short-lived proteins and the regulation of cell cycle and cell growth. The ubiquitin-proteasome pathway is altered in multiple types of human malignancies, including colorectal cancer (CRC). The alteration in the expression of the novel human gene ubiquitin-conjugating enzyme E2 Q1 (UBE2Q1), as a putative member of the E2 ubiquitin-conjugating enzyme family, has been reported in several malignancies, including carcinoma of the breast, hepatocellular and colorectal cancer, and pediatric acute lymphoblastic leukemia. In the present study, the effect of UBE2Q1 overexpression on cell growth, clonogenicity, motility and cell cycle was investigated in a CRC cell line. The UBE2Q1 gene was cloned in the pCMV6-AN-GFP expression vector. A series of stable transfectants of SW1116 cells overexpressing UBE2Q1 protein were established and confirmed by fluorescence microscopy and western blotting. Using these cells, MTT assay was performed to evaluate cell growth and proliferation, while crystal violet staining was used for clonogenicity assay. Cell cycle analysis was also performed to survey the ratio of cells accumulated in different phases of the cell cycle upon transfection. The motility of these cells was also studied using wound healing assay. UBE2Q1 transfectants exhibited a faster growth in cell culture, increased colony formation capacity and enhanced motility compared with control non-transfected cells and cells transfected with empty vector (mock-transfected cells). UBE2Q1 overexpression also resulted in a significant decrease in the number of cells accumulated in the G0/G1 phase of the cell cycle. The present findings suggest that UBE2Q1 may function as an oncogene that induces proliferation of cancer cells, and could be a novel diagnostic tool and a potential therapeutic target for CRC.

Objective Liver cancer is common worldwide and its occurrence is increasing globally; the most prevalent type is hepatocellular carcinoma. Cressa cretica is a halophyte plant that contains flavonoids, which are antibacterial, antioxidant agents. We investigated the effects of C. cretica extract on apoptosis and expression of Bcl2, Bax and P53 genes in the hepatoma cell line, HepG2. Methods HepG2 cells were cultured in RPMI 1640 medium, then incubated in various concentrations of hydroalcoholic extract of C. cretica. Cell proliferation was investigated using MTT assay and apoptosis was measured using flow cytometry. The expression of apoptosis- related genes were determined by real-time PCR. Results Proliferation of HepG2 cells treated with different concentrations of C. cretica were diminished significantly in a dose-dependent pattern. The concentration of C. cretica required for 50% cell viability was 2300 μg/ml. Expression of P53 increased significantly in the treatment groups compared to control group. Bcl2 gene expression was decreased significantly compared to the control group, while the Bax gene was overexpressed in the extract treated groups. Conclusion C. cretica is able to effect on the expression of apoptosis-related genes; therefore, it could play a role in treatment of cancer.

Objective Dyslipidemia is a disorder in lipoprotein metabolism. This disorder could lead to atherosclerosis and it promotes the coronary artery disease and stroke. Turmeric and its active compound, curcumin, modulate the lipid metabolism and also have been used to treat the fatty liver. The present study evaluated the effect of Curcuma longa extract and curcumin on gene expression profiles of lipid metabolism pathway in human HepG2. Methods HepG2 cell line was cultured in RpMI-1640 media. The cells in each well were incubated in various concentration of curcumin and hydro-alcoholic extract of Turmeric (15, 30, 60, 125, 250, 1000, 2000 μg/ml). The cell viability was investigated by MTT assay and the expression of lipid metabolism genes was determined using real-time PCR in IC20 concentration. Results The proliferation of HepG2 cells significantly reduced in curcumin-treated and hydro-alcoholic extract of Turmeric-treated groups with increasing the concentrations. The expression of lipogenesis-related genes such as PGC1A, CPT1A and ACOX1 significantly increase in both treatment groups as compared with the control group. Curcumin and hydro-alcoholic extract of Turmeric significantly diminished the expression of lipid synthesis genes such as SCD1, SREBF2, and DGAT1. Conclusion According to our study, Turmeric and curcumin reduce the viability of HepG2 cells in a concentration-dependent manner; in addition, these ingredients decrease the lipogenesis genes while they increase the lipid synthesis genes. This report suggests that Turmeric could remove the lipid and fatty acids. This herbal substance could be a helpful treatment for obesity and fatty liver.

Objectives: In the present study, we aimed to identify the anti-proliferative potential of [Cu(L)(2imi)] complex [L = 2-(((5-chloro-2-oxyphenyl)imino)methyl)phenolato) and 2imi = 2-methyl imidazole] against HepG2 cells as an in vitro model of human hepatocellular carcinoma and normal mouse fibroblast L929 cells. Methods: The cytotoxic and apoptotic effects of [Cu(L)(2imi)] complex on HepG2 cells and normal fibroblasts (L929) were examined by MTT assay and flow cytometry, respectively. Results: Cytotoxicity induced by [Cu(L)(2imi)] complex was time dependent. Also, there was a positive correlation between cytotoxicity and an increase in Cu complex concentration. For HepG2 cells, the cell viability percentage was 50% at 58 μg/mL after 24 h treatment, whereas in the same concentration and conditions, the viability percentage was surprisingly higher (about 100%) for L929 cells. Also, after 48 h treatment, the viability percentage of HepG2 cells at 55 μg/mL concentration was 50% in contrast with 89.3% for L929 cells in the same conditions. Flow cytometry findings suggest that [Cu(L)(2imi)] complex is capable of decreasing cancer cell viability through apoptosis and did not efficiently activate the necrosis process. Conclusions: Finally, we found that [Cu(L)(2imi)] complex possess the potential for development as an anti-cancer drug for human hepatocellular carcinoma.

Aim of study Diabetes mellitus is related to the development of neuronal tissue injury in different peripheral and central nervous system regions. A common complication of diabetes is painful diabetic peripheral neuropathy (PDN). We have studied the neuroprotective and anti-nociceptive properties of neuropeptide orexin-A in an animal experimental model of diabetic neuropathy. Methods All experiments were carried out on male Wistar rats (220-250 g). Diabetes was induced by a single intraperitoneal injection of 55 mg/kg (i.p.) streptozotocin (STZ). Orexin-A was chronically administrated into the implanted intrathecal catheter (0.6, 2.5 and 5 nM/L, daily, 4 weeks). The tail-flick and rotarod treadmill tests were used to evaluate the nociceptive threshold and motor coordination of these diabetic rats, respectively. Cleaved caspase-3, Bax, Bcl2 and the Bax/Bcl-2 ratio, as the biochemical indicators of apoptosis, were investigated in the dorsal half of the lumbar spinal cord tissue by western blotting method. Results Treatment of the diabetic rats with orexin-A (5 nM/L) significantly attenuated the hyperalgesia and motor deficit in diabetic animals. Furthermore, orexin-A (5 nM/L) administration suppressed pro-apoptotic cleaved caspase-3 and Bax proteins. Also, orexin-A (5 nM/L) reduced the expression of Bax/Bcl-2 ratio in spinal cord dorsal half of rats with PDN. Conclusions Altogether our data suggest that the orexin-A has anti-hyperalgesic and neuroprotective effects in rats with PDN. Cellular mechanisms underlying the observed effects may, at least partially, be related to reducing the neuronal apoptosis.

Background: Cancer is one of the major threatening factors of human health worldwide. Unfortunately, chemotherapy, the powerful arm of cancer therapy, is accompanied with many side effects, so alternative treatments with greater specificities and fewer side effects are highly required. Methods: Human cancer cell lines including SW-742, HCT116, HepG2, Hep2, MKN45 and LNcap were selected and the anti-cancer potential of Cyrtopodion scabrum extract (CsE) on their growth was studied. Vero cells were used to study the potential cytotoxicity on the normal cells. Cell cycle analysis and DNA fragmentation assay were also performed. Results: CsE was toxic (30% - 78%) to all the cell lines, with the highest cytotoxicity on SW742, MKN45 and HepG2, respectively. A high selectivity index (> 2) was observed for the extract on SW742 and MKN45 cell lines. DNA laddering pattern, as well as a significant increase in the number of the cells accumulated in sub-G1 and G2-phase of the cell cycle compared to the control untreated cells, was also observed. Conclusions: CsE suppressed the human cancer cells selectively and probably through apoptosis and G2 arrest mechanism. It could suggest a promising alternative/complementary treatment for cancer patients, especially those who suffer from digestive tract cancer.

Objective Hyperglycemia in patients with Diabetic Mellitus (DM) results from the demolition of islet β cells in the pancreas as well as failure in insulin action or poor response to insulin secretion and there are no further methods to treat diabetes apart from insulin injection and oral hypoglycemic medicines. Thus, the main objective of the current research investigates hydro-alcoholic extracts of Prosopis farcta on impact on the Pyruvate kinase (Pk), Phospho fructo kinase (PFK), Gluco kinase (GK) and signaling insulin Phosphotidylinositol 3-Kinase (PI3K), Insulin Receptor Substrate (IRS1), Glucose Transporter type2 (GLUT2) in HepG2 cells. Method By various concentrations (0–10 mg/ml) of the extract (leaf-root – fruit) for 24 and 48 h, HepG2 cells were cultured and treated in this experimental research. MTT test evaluated cell proliferation by ELISA Reader measured the optical density of the colored solution at 570 nm wavelengths. Real-time-PCR method defined the fold changes of genes expression against β-actin. Data were analyzed by ANOVA. Variations with a P-value Results Our results showed cells were treated with various concentrations of extract (leaf-root – fruit) for 24 and 48 h had markedly declined cell viability in HepG2 cell line as compared to controls in a time and dose-dependent methods, an evident deviation is seen between different concentrations of the extracts. Real Time-PCR analysis showed that the expression of glucokinase, phosphofructo and pyruvate kinase were down regulated and expression of IRS1-PI3K-GLUT2 genes were up regulated in compare to control cells (P Conclusion It appears the anti-diabetic impact of the Prosopis farcta plant not to be over the glycolysis path. This acts mechanisms via enhancing the gene expression in the insulin signaling pathway. The plant, thus, may serve to enhance the insulin secretion function through triggering insulin signaling paths and, in turn, inhibiting increased blood glucose impact on diabetic patients. It is, therefore, expected that the various extracts of such plant may have an impact on diabetic patients, type-2 diabetes in particular.

Inflammatory bowel disease (IBD) is a disease strongly associated with colorectal cancer (CRC) as a well-known precancerous condition. Alterations in DNA methylation and mutation in K-ras are believed to play an early etiopathogenic role in CRC and may also an initiating event through deregulation of molecular signaling. Epigenetic silencing of APC and SFRP2 in the WNT signaling pathway may also be involved in IBD-CRC. The role of aberrant DNA methylation in precancerous state of colorectal cancer (CRC) is under intensive investigation worldwide. The aim of this study was to investigate the status of promoter methylation of MGMT-B, APC1A and SFRP2 genes, in inflamed and normal colon tissues of patients with IBD compared with control normal tissues. A total of 52 IBD tissues as well as corresponding normal tissues and 30 samples from healthy participants were obtained. We determined promoter methylation status of MGMT-B, SFRP2 and APC1A genes by chemical treatment with sodium bisulfite and subsequent MSP. The most frequently methylated locus was MGMT-B (71%; 34 of 48), followed by SFRP2 (66.6 %; 32 of 48), and APC1A (43.7%; 21 of 48). Our study demonstrated for the first time that hypermethylation of the MGMT-B and the SFRP2 gene promoter regions might be involved in IBD development. Methylation of MGMT-B and SFRP2 in IBD patients may provide a method for early detection of IBD-associated neoplasia.

Breast and colorectal cancers rank high in Iran as causes of mortality. Most of the current treatments are expensive and non-specific. The potential anticancer properties of common home gecko, Cyrtopodion scabrum, were investigated in this study. The effects of C. scabrum extract on proliferation, viability and migration of the colorectal cancer (SW-742), breast cancer (MCF-7) and normal (MSC) cell lines were investigated using MTT and in vitro wound healing assay. IC50 values calculated for the extract were 559±28.9 μg/ml for MCF-7 and 339±11.3 μg/ml for SW-742. No toxic effects on the normal control cells were observed. MCF-7 and SW-742 cell growth was inhibited by 32.6% and 62%, under optimum conditions, compared to the untreated control cells. The extract also decreased the motility and migration ability of both cancer cell lines, with no significant effects on the normal control cells. Data suggest C. scabrum extract as a useful natural resource for targeting cancer cells specifically.

BackgroundOur objective was to determine the role of Interleukin-6 (IL-6) and Tumor Necrosis Factor-alpha (TNF-alpha), markers of immune activation and endothelial dysfunction, in patients with preeclampsia.MethodsTwenty four women with preeclampsia and eighteen antepartum normotensive pregnant women were recruited as controls. Serum levels of IL-6 and TNF-alpha were measured by enzyme-linked immunosorbent assay. We used independent-samples t test to assess the differences in the concentration of cytokines in preeclamptic patients and control subjects.ResultsIL-6 levels [mean (S.D.)] were significantly higher in preeclamptic women [5.8 (4.85) pg/ml] compared to normal pregnant women [3.01 (2.45) pg/ml] (p = 0.02). There was no significant change in concentration of TNF-alpha in preeclamptic women [53.8 (30.0) pg/ml] compared to normal pregnant women [51.9 (33.8) pg/ml] (p > 0.1).ConclusionThe results of this study show that IL-6 as a pro-inflammatory cytokine is present in higher concentration in women with preeclampsia. The study was undertaken in women with established preeclampsia and it is not possible to determine whether the increased concentration of IL-6 is a cause or consequence of the disease. Furthermore, these findings suggest that serum TNF-alpha level is not associated with preeclampsia.