Crocin Suppresses Colorectal Cancer Cell Proliferation by Regulating miR-143/145 and KRAS/RREB1 Pathways.

BACKGROUND: As a chemoprevention agent, crocin effectively decreases the risk of human cancers, including colorectal cancer (CRC). However, the mechanism underlying the anti-cancer effects of crocin is not entirely explained. Considering that in this study, we investigated the crocin effect on miR-143/145 and related signaling pathways in CRC cells. METHODS: HCT-116 and HT-29 CRC cells were treated with different concentrations of crocin and then were subjected to MTT and qRT-PCR assays to investigate cell viability and miR-143/miR-145, KRAS, and RREB1 expression, respectively. Also, western blotting was performed to evaluate gene expression at protein levels. RESULTS: Our results showed that treating CRC cells with crocin decreases cell viability by upregulating miR-143/145 expression and reducing KRAS and RREB1 expression dose-dependently. These effects on gene expression in CRC cells were reversed by removing crocin from the media after 48 h. Furthermore, western blotting results exhibited that crocin significantly reduced the protein expression of KRAS and RREB1. Also, it was found that treatment of CRC cells by crocin led to the inactivation of AKT by decreasing its phosphorylation. CONCLUSIONS: This study suggests that crocin may inhibit CRC cell proliferation by modulating KRAS, REEB1, and AKT signaling pathways mediated through miR-143/145 upregulation.

Silencing of SiX-4 enhances the chemosensitivity of melanoma cells to Cisplatin.

Melanoma is the riskiest type of skin cancer. Its prevalence has been rapidly increased over the last three decades. SIX1, SIX2, SIX3, SIX4, SIX5, and SIX6 are members of the sine oculis homeobox (SIX) homolog family. It is imperative to identify new melanoma biomarkers to improve the predictive value for melanoma prognosis, which could enhance our understanding of carcinogenesis and tumor progression. In this study, we investigated whether silencing of SIX4 in a melanoma cell line (A375 cells) in combination with Cisplatin can affect the apoptosis and suppression of cell cycle progression, migration of the melanoma cells. MTT test and colony formation assay was applied to determine the IC50 of Cisplatin and the combined effect of SIX4 siRNA and Cisplatin on the viability and clonogenesis of the A-375 cells. qRT-PCR was performed to determine the c-myc, BCL-2, BAX, MMP-9, CXCR4, and Rock genes expression. Furthermore, flow cytometry was applied to evaluate apoptosis, autophagy, and the cell cycle status in different groups. Finally, wound healing assay was employed to evaluate the effect of this combination therapy on migratory capacity. SIX4 suppression increased the chemosensitivity of A-375 cells to Cisplatin and decreased its efficient dose. Furthermore, SIX4 suppression alongside Cisplatin reduced cell migration rate, arrested the cell cycle at the G1 phase, induced apoptosis by modulating the expression of apoptotic target genes, induced autophagy, and also significantly inhibits clonogenesis of A-375 cells. SIX4 plays a significant role in the chemosensitivity and pathogenesis of melanoma. Therefore, SIX4 suppression, in combination with Cisplatin, may be a promising therapeutic approach in treating melanoma.

Ultrasensitive bioassay of epitope of Mucin-16 protein (CA 125) in human plasma samples using a novel immunoassay based on silver conductive nano-ink: A new platform in early stage diagnosis of ovarian cancer and efficient management.

Ovarian cancer (OC) is known to be one of the most lethal malignancies associated with women disease. The CA-125 protein is a repetitive epitope of MUC-16, which plays key role in enhancing the proliferation of cancer cells and inhibiting anticancer immune responses. It is the most widely used biomarker for early stage diagnosis of OC. Also it is the only serum marker which currently used in clinical diagnosis. Monitoring of CA-125 protein in the serum sample is also valuable in evaluating the response of ovarian cancer to treatment. In this research, a novel immunoassay based on immobilization of CA-125 antibody on the biointerface of silver nanoparticles modified graphene quantum dots ink (Ag NPs-GQDs) was successfully designed to recognition of CA-125 protein in a human plasma sample. The supplied immunoassay presents the proper ability to detect and determine the amount of CA-125 biomarker in low concentrations of CA-125 biomarker. The proposed immunosensor was employed for the detection of CA-125 using differential pulse voltammetry (DPVs) and square wave voltammetry (SWVs) techniques. The proposed interface leads to enhancement of accessible surface area for immobilizing a high amount of anti-CA-125 antibody, increasing electrical conductivity, boosting stability, catalytic properties and biocompatibility. Under the optimized operating conditions, the low limit of quantitation (LLOQ) for the proposed immunosensor was recorded as 0.01 U/mL, which this evaluation was performed at highly linear range of 0.01-400 U/mL. The proposed immunoassay was successfully applied for the monitoring of CA-125 in unprocessed human plasma samples.

Ultrasensitive immunoassay of tumor protein CA 15.3 in MCF-7 breast cancer cell lysates and unprocessed human plasma using gold nanoparticles doped on the structure of mesoporous silica.

Breast cancer is the most common threat in women worldwide. Increasing death rate of diagnosed cases is the main leading cause of designing specific immunoassay for tumor marker in breast cancer. Cancer antigen 15-3 (CA 15-3) is a tumor protein for many types of cancer, most notably breast cancer. Herein, we report a sandwich-type electrochemical immunosensor based on signal amplification strategy of multiple nanocomposites to detection of CA 15-3 biomarker. The proposed immunosensor fabricated by reduced graphene oxide (rGO), poly-dopamine (PDA) and amino functionalized mesoporous silica (MCM-41-NH2) doped by gold nanoparticles composite on the glassy carbon electrode with a large surface area which was prepared a new platform to immobilization of primary antibodies and provide excellent conductivity. To further amplify the electrochemical signal, the trace tag on the foundation of gold nanoparticles (AuNPs) is coated with MCM-41-NH2 nanocomposite through thionine linking, which provides more amino groups to capture more horseradish peroxidase-labeled antibodies (HPR-Ab2) and enhances the conductivity. Under optimal conditions, the developed immunosensor exhibits excellent analytical performance for the determination of CA 15-3 with a wide linear range from 0.002 to 125 U/mL and a low limit of quantification of 0.002 U/mL. Furthermore, satisfactory results are obtained for the determination of CA 15-3 in human plasma samples, indicating the potential of the immunoassay to be applied in clinical analysis.

Ultrasensitive immunoassay of carcinoma antigen 125 in untreated human plasma samples using gold nanoparticles with flower like morphology: A new platform in early stage diagnosis of ovarian cancer and efficient management.

Ovarian cancer, as one of the most life-threatening malignancies among women worldwide, is usually diagnosed at the late stage despite up regulation of molecular markers such as carcinoma antigen 125 (CA 125) at the early stages of the malignancy. CA 125 is the only tumor marker recommended for clinical use in the diagnosis and management of ovarian cancer. The potential role of CA-125 for the early detection of ovarian cancer is controversial and has not yet been adopted for widespread screening efforts in asymptomatic women. Therefore, early detection of CA 125 in human biofluids is highly demanded. In the present study, a novel method was proposed for the fabrication of electrochemical immunosensor based reduced graphene oxide (RGO). Cysteamine capped gold nanoparticle (Cys-AuNPs) were deposited over the surface of ERGO probe using electrophoretic deposition method. These Cys-AuNPs/ERGO probes provide the favorable sites to attach the monoclonal antibody specific to CA 125 antigen. Cyclic voltammetry (CV), and square wave voltammetry (SWV) were applied for the electrochemical recognition of the biolayer. The represented signals demonstrates excellent figure of merits and good capability of the engineered immunosensor towards sensitive detection of CA 125. Quantitative measurements of CA 125 in human plasma samples have been demonstrated, showing the potential of the practical application of this novel immunosensor for the analysis of this biomarker in blood serum samples. This immunosensor has the ability of direct electron transfer as compared to earlier reported electrochemical immunosensors based electrochemical methods. Further, this immunosensor provides a very suitable and convenient alternative to replace the expensive commercially available methods such as immunohistochemistry. The following regression equation between the electrochemical current response and the CA 125 concentration range from 0.1 to 400 U/mL was found. The low limit of quantification for this immunosensor was 0.1 U/mL. To the best of our knowledge, this is the first reported on the direct immobilization of antibody on the surface of Cys-AuNPs/ERGO for fabrication of immunosensors.

An innovative immunosensor for ultrasensitive detection of breast cancer specific carbohydrate (CA 15-3) in unprocessed human plasma and MCF-7 breast cancer cell lysates using gold nanospear electrochemically assembled onto thiolated graphene quantum dots.

The accurate quantification of the level of breast cancer specific protein CA 15-3 in serum is crucial for cancer prognosis. This work, a novel and sensitive label-free immunoassay based on gold nanospear (Au NSs) electrochemically assembled onto thiolated graphene quantum dots (CysA/GQDs) for the detection of CA 15-3 antibodies. The CysA/Au NSs/GQDs hybrid interface provides a large surface area for the effective immobilization of CA 15-3 antigens, as well as it ascertains the bioactivity and stability of immobilized CA 15-3 antigens. Field emission scanning electron microscope (FE-SEM), and EDS photoelectron spectroscopies were used to monitor the sensor fabrication. Also, cyclic voltammetry was used to quantify the extent of Au NSs' surface coverage by CA 15-3 antigens. Square wave voltammetry (SWV) was employed to investigate the immunosensor fabrication and to monitor the binding events between CA 15-3 antigens-antibodies. Under optimized experimental conditions, the immunosensor displayed good sensitivity and specificity. The CA 15-3 were detected in a concentration as low as 0.11U/mL with a linear range from 0.16-125U/mL. The high sensitivity of the immunosensor may derive from the high loading of CA 15-3 antibodies on CysA/Au NSs/GQDs hybrid interface which increases the number of binding events. The method was successfully applied assay of the CA 15-3 in unprocessed human plasma samples. Also, proposed immunosensor was applied to the assay of CA 15-3 malignant cell line lysates (human breast adenocarcinoma cell line-MCF-7).

Angiogenic potential of YKL-40 in the dynamics of tumor niche.

A multitude of clinical studies showed the elevation of YKL-40 in subjects with different kinds of tumors. It is predicted that an inherent correlation exists between survivals of cancer patients with total YKL-40 serum levels, making this factor as a potential novel biomarker. However, the crucial role of YKL-40 in the dynamics of cancers, especially angiogenesis, has not yet been completely addressed. In this review, we highlighted the various facets of YKL-40 and its importance in cancer biology as a bio-shuttle in gene therapy.

Aptamer based assay of plated-derived grow factor in unprocessed human plasma sample and MCF-7 breast cancer cell lysates using gold nanoparticle supported α-cyclodextrin.

Platelet-derived growth factor (PDGF), a protein biomarker, is directly involved in many cell transformation processes, such as tumor growth and progression. Elevation platelet-derived growth factor (PDGF-BB) concentration in plasma could indicate the accelerating growth of metastatic breast tumors and angiogenesis. The development of an apta-assay for detection of PDGF-BB in is presented in this work. A highly specific DNA-aptamer, selected to PDGF-BB was immobilized onto a gold nanoparticles supported α-cyclodextrin and electrochemical measurements were performed in a solution containing the phosphate buffer solution with physiological pH. Variety of shapes of gold nanostructures with different sizes from zero-dimensional nanoparticles to spherical structures were prepared by one-step template (α-cyclodextrin)-assistant green electrodeposition method. Fully electrochemical methodology was used to prepare a new transducer on a gold surface which provided a high surface area to immobilize a high amount of the aptamer. The surface morphology of electrode was characterized by high-resolution field emission scanning electron microscope (FE-SEM) and energy dispersive spectroscopy (EDX). The prepared aptasensors represented different electrochemical activities toward the redox processes of PDGF-BB attributing to the size and shape of the gold nanoparticles. The aptasensor was employed for the detection of PDGF using square wave voltammetry (SWV) and Cyclic voltammetry (CV) techniques. Under optimized condition the calibration curve for PDGF-BB was linear in 0.52-1.52nM with low limit of quantification of 0.52nM. Also, under the optimized experimental conditions, the proposed aptasensor of GNPs-cubic-α-CD-Apt-Au electrode exhibited excellent analytical performance for MCF-7 cells determination, ranging from 328 TO 593 cells mL-1 with low limit of quantification of 328 cells mL-1. As a result, the electrochemical aptasensor was able to detect cancer-related targets in unprocessed human plasma samples.

An innovative immunosensor for detection of tumor suppressor protein p53 in unprocessed human plasma and cancer cell lysates.

An innovative mediator-free electrochemical immunosensor for quantitation of p53 tumor suppressor protein based on signal amplification strategy was fabricated. In this work, biotin conjugated p53-antibody (anti-p53) was immobilized onto a green and biocompatible nanocomposite containing poly l-cysteine (P-Cys) as conductive matrix and 3D gold nanoparticles (GNPs) as signal amplification element. Therefore, a novel nanocomposite film based on P-Cys and GNPs was exploited to develop a highly sensitive immunosensor for detection of p53 protein. Importantly, GNPs prepared by sonoelectrodeposition method which lead to compact morphology. Fully electrochemical methodology was used to prepare a new transducer on a gold surface which provided a high surface area to immobilize a high amount of the anti-p53. The surface morphology of electrode was characterized by high-resolution field emission scanning electron microscope (FE-SEM) and energy dispersive spectroscopy (EDX). The immunosensor was employed for the detection of p53 in physiological pH using square wav voltammetry and differential pulse voltammetry (DPVs) techniques. Under optimized condition the calibration curve for p53 concentration by SWV and DPV was linear in 0.0369-50pM and 0.018-2.5pM with lower limit of quantification of 48fM and 18fM, respectively. The method was successfully applied assay of the p53 in unprocessed human plasma samples. Also, the method was applied to the assay of p53 in human plasma sample and normal and malignant cell line lysates such as (L929 normal cell Line from mouse C3H (L929), colon cancer cell-HCT, prostate cancer cell line PC-3, and human breast adenocarcinoma cell line-MCF7).

Poly arginine-graphene quantum dots as a biocompatible and non-toxic nanocomposite: Layer-by-layer electrochemical preparation, characterization and non-invasive malondialdehyde sensory application in exhaled breath condensate.

This study reports on the electropolymerization of a low toxic and biocompatible polymer with entitle poly arginine-graphene quantum dots (PARG-GQDs) as a novel strategy for surface modification of glassy carbon (GC) surface and preparation a new interface for biomedical application. The fabrication of PARG-GQDs on GCE was performed using Layer-by-layer regime. Scanning electron microscopy (SEM) was confirmed dispersion of GQDs on the surface of PARG which lead to increase of surface coverage of PARG. The redox behavior of prepared sensor was then characterized by cyclic voltammetry (CV), differential pulse voltammetry (DPV) and chronoamperometry (CHA), square wave voltammetry (SWV), linear sweep voltammetry (LSV). The electroactivity of PARG-GQDs coating towards detection and determination of malondialdehyde (MDA) as one of the most common biomarkers of oxidative stress, was then studied. Then, application of prepared sensor for the detection of MDA in exhaled breath condensate (EBC) is described. Electrochemical based sensor shows the lower limit of quantification (LLOQ) were 0.329nanomolar. This work is the first report on the integration of GQDs to poly amino acids. Further development can lead to monitoring of MDA or other exhaled breath biomarkers by GQDs functionalized poly amino acids in EBC using electrochemical methods.

Poly-dopamine-beta-cyclodextrin: A novel nanobiopolymer towards sensing of some amino acids at physiological pH.

A novel nanobiopolymer film was electrodeposited on the surface of glassy carbon through cyclic voltammetry from dopamine, ß-cyclodextrin, and phosphate buffer solution in physiological pH (7.40). The electrochemical behavior of polydopamine-Beta-cyclodextrin modified glassy carbon electrode was investigated for electro-oxidation and determination of some amino acids (l-Cysteine, l-Tyrosine, l-Glycine, and l-Phenylalanine). The modified electrode was applied for selected amino acid detection at physiological pH using cyclic voltammetry, differential pulse voltammetry and chronoamperometry, chronocoulometery. The linear concentration range of the proposed sensor for the l-Glycine, l-Cysteine, l-Tyrosine, and l-Phenylalanine were 0.2-70, 0.06-0.2, 0.01-0.1, and 0.2-10µM, while low limit of quantifications were 0.2, 0.06, 0.01, and 0.2µM, respectively. The modified electrode shows many advantages as an amino acid sensor such as simple preparation method without using any specific electron transfer mediator or specific reagent, good sensitivity, short response time, and long term stability.

The Effect of Tomato Juice Consumption on Antioxidant Status in Overweight and Obese Females.

Tomatoes and their products are the main source of lycopene, a powerful potent antioxidant. Tomato products improve antioxidant defenses and reduce the risk of oxidative stress, at least partly, due to the presence of lycopene. Lycopene, as an antioxidant, induces the upregulation of antioxidant enzymes and reinforces the total enzyme capacity of the human body. Obesity is a chronic condition in which destructive mechanisms increase the reactive oxygen species and attenuation of antioxidant status. We hypothesized that the consumption of a lycopene-rich food would improve the antioxidant defense of women who were overweight or obese. A total of seventy-five overweight or obese female students of the Tehran University of Medical Sciences were enrolled and randomly allocated to one of two groups, intervention (n = 40) or control (n = 35), consuming 330 ml/d of tomato juice or water, respectively, for a 20-day period. At baseline and day 20, total antioxidant capacity and antioxidant enzyme activity (superoxide dismutase, glutathione peroxidase, and catalase) were analyzed using ELISA kits and spectrophotometric methods and then compared between the two groups. Lycopene consumption had no effect on these aforementioned variables. Therefore, it seems that more research with longer duration and more sensitive indicators will be required.

Protective effect of soy protein on collagen-induced arthritis in rat.

To evaluate preventive and therapeutic effects of soy protein on collagen-induced arthritis rats. Sprague-Dawley rats immunized with bovine type II collagen emulsified in adjuvant and treated with soy protein (7 g/kg), dexamethasone (1 mg/kg), and casein (in control groups) by daily gavages feedings for 30 days. Score of arthritis recorded every day for each paws of animal. Tumor necrosis factor-alpha, interleukin6, leptin, and adiponectin were measured in serums. Treatment with soy protein resulted in significant delay in time to onset of arthritis as well as significantly decreased arthritis incidence, clinical arthritis severity score, histopathological arthritis severity score, and in vivo cell-mediated immunity to collagen (P < 0.05). Administration of soy protein significantly suppressed the progression of collagen II-induced arthritis and inhibited the production of tumor necrosis factor-alpha, interleukin6, leptin, and adiponectin. Soy protein appeared to be a potent immunomodulatory inhibitor of collagen II-induced arthritis in rats. It could delay onset of RA and reduced cartilage erosion and synovitis inflammation. Therefore, it may be a useful protein in the prevention and treatment of rheumatoid arthritis patient.

Sensitivity and specificity of a short questionnaire for food insecurity surveillance in Iran.

BACKGROUND: Food insecurity is frequent in both developed and developing countries, affecting from 5% to 25% of the general population. It has considerable health impacts on the physical, social, and psychological status of individuals in communities suffering from food insecurity. OBJECTIVE: The aim of this study was to document the epidemiologic features of food insecurity in the northwest region of Iran and to evaluate the sensitivity and specificity of a short-form (six items) questionnaire for screening of food insecurity in the region. METHODS: This cross-sectional study was conducted on 300 subjects (132 male and 168 female) selected randomly in the Asadabadi area of the northwest of Iran. Information on food consumption was obtained by a 24-hour food-recall questionnaire for 3 days in a week. This information was compared with the data from the Household Food Security Scale (six-item short questionnaire) to assess the applicability of this short scale for the surveillance of food insecurity. Hunger was defined as inadequate intake of energy. Hidden hunger was defined as adequate intake of energy and inadequate intake of one (or more) of four key nutrients (protein, calcium, vitamin A, and vitamin B2). RESULTS: The prevalence of hunger and hidden hunger in the area according to the 24-hour food-recall questionnaire was 26% and 42%, respectively. Only 32% of the study population was secure in terms of having access to all key nutrients. The sensitivity, specificity, and accuracy of the short questionnaire for screening for hunger in the population were 98.7%, 85.5%, and 89%, respectively; and the corresponding values for hidden hunger were 23.5%, 96.9%, and 56.3%. CONCLUSIONS: Our findings indicate that food insecurity is prevalent in the northwest of Iran. The short questionnaire (six items) may be used as a simple, low-cost, rapid, and useful tool for the screening of food insecurity and energy intake in similar areas.

Effect of retinol on iron bioavailability from Iranian bread in a Caco-2 cell culture model.

OBJECTIVE: Iron and vitamin A deficiencies are among the most prevalent nutritional problems. There are no data on iron bioavailability in Iran. In addition, interaction of vitamin A with iron bioavailability is not well documented, so we determined iron bioavailability from the most widely consumed food in Iran, lavash bread, and the effect of vitamin A on this bioavailability. METHODS: In vivo human studies for determining iron bioavailability are cumbersome, time consuming, and costly to perform, so we used an in vitro model in Caco-2 cells. Bread samples were digested with or without vitamin A (10 microg/1.0 g of dried bread). We used an iron solution containing vitamin C as a positive control. Iron absorption was measured using 59FeCl3. Bioavailability was defined as the percentage of radiolabeled iron taken up and transferred by Caco-2 cells after 1.5 h of incubation. Experiments were carried out two to four times. RESULTS: Mean +/- standard deviations for iron bioavailability in bread samples digested without or with additional vitamin A and in controls were 2.53 +/- 1.55%, 6.62 +/- 3.40%, and 20.80 +/- 2.30%, respectively (P < 0.001). Vitamin A caused a 2.62-fold increase in iron absorption from bread samples. CONCLUSIONS: Iranian lavash bread has low iron bioavailability, but this can be increased by vitamin A supplementation. Increasing vitamin A intake can be considered as a method for increasing iron bioavailability, thus combating iron and vitamin A deficiencies simultaneously.

Development and validation of a short food-frequency questionnaire for screening women of childbearing age for vitamin A status in northwestern Iran.

A high percentage of women in their childbearing years suffer from subclinical vitamin A deficiency; 10% to 20% of pregnant women worldwide are vitamin A deficient. This study aimed to design and validate a short food-frequency questionnaire to serve as a simple screening tool for vitamin A status in women of childbearing age. The sample consisted of 187 healthy, nonpregnant, nonlactating women 15 to 49 years of age, from urban and rural areas of Marand district in East Azerbaijan. Dietary intake was evaluated by a face-to-face interview using a 24-hour dietary recall for two consecutive days and a 41-item qualitative food frequency questionnaire. Height, weight, and serum retinol were measured. Serum retinol values were less than 20 micrograms/dl for three subjects, while an additional 34 subjects (18%) had values between 21 and 30 micrograms/dl. Principal-component analysis performed on the food-frequency questionnaire identified five components that together defined 34.4% of the variance in estimated vitamin A intake and were used to derive a 20-item short food-frequency questionnaire. Internal consistency of the short instrument was acceptable (Cronbach's alpha = .59). Serum retinol was significantly correlated with total vitamin A intake and with intake of vitamin A from plant sources, as estimated by the short food-frequency questionnaire. Important sources of provitamin A in these women's diets included some not typical of other populations: nuts and green leaves of types used elsewhere in small quantities as herbs, but important in Iran because the amount and frequency of consumption are relatively high. We conclude that the questionnaire is relatively valid and potentially useful in identifying women at risk for vitamin A deficiency in this population.