Evaluation of tumor regulatory genes and apoptotic pathways in the cytotoxic effect of cytochalasin H on malignant human glioma cell line [U87MG]

Objective:The aim of current study was to provide a proof-of-concept on the mechanism of PLAU and PCDH10 gene expressions and caspases-3, -8, and -9 activities in the apoptotic pathway after treatment of malignant human glioma cell line [U87MG] with cytochalasin H

Materials and Methods: In the present experimental study, we have examined cytochalasin H cytotoxic activities as a new therapeutic agent on U87MG cells in vitro for the first time. The cells were cultured and treated with 10-5-10-9M of cytochalasin H for 24, 48 and 72 hours. The assessment of cell viability was carried out by [3-[4,5-dimethylthiazol-2-yl]- 2,5-diphenyltetrazoliumbromide [MTT] assay at 578 nm. The data are the average of three independent tests. mRNA expression changes of PLAU and PCDH10 were then evaluated by quantitative reverse-transcriptase polymerase chain reaction [qRT-PCR]. The fluorometric of caspases-3, -8, and -9 activities were carried out. The morphology changes in the U87MG cells were observed by fluorescence microscope

Results: MTT assay showed that cytochalasin H [10-5 M] inhibited the U87MG cancer cells proliferation after 48 hours. Analysis of qRT-PCR showed that the PLAU expression was significantly decreased in comparison with the control [P<0.05]. The expression of PCDH10 also showed a significant increase when compared to the control [P<0.001]. Fluorescence microscope indicated morphological changes due to apoptosis in U87MG cancer cells, after treatment with cytochalasin H [10-5M, 48 hours]. The fluorometric evaluation of caspase-3, -8, and -9 activities showed no significant difference between the caspases and the control group

Conclusion: This study shows the effect of caspase-independent pathways of the programmed cell death on the U87MG cancer cell line under cytochalasin H treatment. Further studies are needed to explore the exact mechanism

Artificial neural network analysis in preclinical breast cancer

In this study, artificial neural network [ANN] analysis of virotherapy in preclinical breast cancer was investigated. In this research article, a multilayer feed-forward neural network trained with an error back-propagation algorithm was incorporated in order to develop a predictive model. The input parameters of the model were virus dose, week and tamoxifen citrate, while tumor weight was included in the output parameter. Two different training algorithms, namely quick propagation [QP] and Levenberg-Marquardt [LM], were used to train ANN. The results showed that the LM algorithm, with 3-9-1 arrangement is more efficient compared to QP. Using LM algorithm, the coefficient of determination [R[2]] between the actual and predicted values was determined as 0.897118 for all data. It can be concluded that this ANN model may provide good ability to predict the biometry information of tumor in preclinical breast cancer virotherapy. The results showed that the LM algorithm employed by Neural Power software gave the better performance compared with the QP and virus dose, and it is more important factor compared to tamoxifen and time [week]

Translation elongation rate measurement of Epstein- Barr virus strain GD1

Epstein-Barr Virus [EBV] has a great co relationship with human malignancies such as gastric carcinoma. Synonymous codon investigations in viruses could help designing vaccine, to generate immunity. Codon Adaptation Index [CAI] has measured translation elongation rate, among the highly expressed genes. The aim of this study was: usage of "CAI" to measure translation efficiency to know how fast EBV-GD1 could produce its proteins. The complete genomic sequences of human herpes virus 4 strain GD1 have retrieved from http://www.ncbi.nlm.nih.gov/sites/gquery [GenBank accession no. AY961628] to extract all protein-coding genes. The sequences have analyzed with DAMBE software. The results have shown that CAI values for the EBV-GD1 genes were 0.76356 +/- 0.02957. The highest and lowest CAI values were 0.82233 and 0.68321 respectively. The results have shown that highly expressed genes mostly had more codon usage bias than low expressed genes. The results provide and introduce not only a system, but also the principles in order to understand the pathogenesis and evolution of EBV-GD1, to open a window, in order to make a better product or vaccine to challenge with the virus

Virotherapy in cancer

New cancer therapies with novel mechanisms and functions are needed to treat patients with different cancers. Virotherapy is a good scenario for such treatment. The advantages of virotherapy include the potential lack of cross resistance with standard therapies and the ability to cause tumor destruction by numerous mechanisms. Oncolytic virus not only possesses unique mechanisms of action that are distinct from other treatment modalities, its self-perpetuating nature provides an ideal platform for therapeutic transgenic insertion. In this review article, a variety of oncolytic viruses in cancer gene therapy will be described

Extract of azadirachta indica [neem] leaf induces apoptosis in 4T1 breast cancer BALB/c mice

Azadirachta indica [Neem] has been used traditionally for many centuries. Some impressive therapeutic qualities have been discovered. However, the therapeutic effect of neem leaf extract in 4T1 breast cancer has not been documented. The purpose of the present study is to investigate the therapeutic effect of ethanolic Neem leaf extract in an in vivo 4T1 breast cancer model in mice. A total of 84 female BALB/c mice were divided randomly into 7 groups [3 non-cancerous groups and 4 cancerous groups] consisting of 12 mice per group. The 3 non-cancerous groups were normal mice treated with 0.5% of Tween 20 in phosphate buffer saline [PBS] [NC], 250 mg/kg Neem [N250] or 500 mg/kg Neem [N500]. The 4 cancerous groups were; cancer controls treated with 0.5% of Tween 20 in PBS [CC], and cancerous mice treated with 0.5 micro g/mL tamoxifen citrate [CT], 250 mg/kg Neem leaf extract [CN 250] or 500 mg/kg Neem leaf extract [CN 500]. Terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL] assays were used to evaluate apoptosis [cell death] in the breast cancer tissues. SPSS software, version 14 was used for statistical analysis. Statistical significance was defined as p<0.05.Non parametric analysis of variance [ANOVA] was performed with the Kruskal Wallis test for the TUNEL assays. Parametric data among the groups was compared using ANOVA. TUNEL assays showed that the CN 250 and CN 500 groups had a higher incidence of apoptosis compared with the cancer controls. The findings showed that neem leaf extract induces apoptosis in 4T1 breast cancer BALB/c mice

Effect of berberis vulgaris aqueous extract on the apoptosis, sodium and potassium in hepatocarcinogenic rats

The effect of Berberis vulgaris aqueous extract in hepatocarcinogenic rats was studied to investigate the apoptotic and sodium, potassium elements properties. A loss of both intracellular potassium and sodium occurs when apoptotic cells shrink and prior to the loss of membrane integrity. Forty-eight Sprague dawley rats were randomly divided into 2 groups, normal and cancerous. Each group was divided into 4 subgroups. The first subgroup acted as normal control while the others were treated with 25, 50 and 100 mg/kg of Berberis vulgaris extract [BVE] and respectively considered as NC, NC25, NC50 and NC100. The first subgroup of cancerous rats acted as cancer control while the others were treated with 25, 50 and 100 mg/kg of BVE and considered as C, C25, C50 and C100. Ion selective electrode [ISE] method was used to measure the level of sodium, potassium, and chloride. TUNEL assay used for the detection of apoptosis cells. Microscopic observations of the TUNEL-positive apoptotic cells revealed a significant difference [P < 0.05] between cancer control [C] and normal control [NC] group. The results indicated that increasing concentration of Berberis vulgaris aqueous extract in cancerous treated groups [C25, C50 and C100] showed an increasing considerabl changes [P < 0.05] of TUNEL-positive cells compared with the cancer control group [C]. Sodium and chloride levels were significantly different [P < 0.05] in cancer control group [C] compared to normal control group [NC]. The results suggested that apoptotic cells level was increased with the BVE concentration in cancerous groups. The Berberis vulgaris extract shows to playa prominent role in promoting apoptosis on the treatment and it is dose dependent