Cytolytic Effects of Newcastle Disease Virus Strain Af2240 on Dbtrg.05mg and U-87mg Brain Tumor Cell Lines

Newcastle disease virus (NDV) is a potential oncolytic agent as it can replicate up to 10,000 times better in human cancer cells than in most normal human cells. Several strains of NDV were reported to induce cytolysis to various cancerous cell lines. In this study, the cytolytic effects of local strain NDV AF2240 toward DBTRG.05MG (glioblastoma multiform) and U-87MG (anaplastic astrocytoma) cell lines were determined using microtetrazolium assay (MTT) for both monolayer and co-culture methods. The value of (IC50) inhibition concentration, fifty percent at which the titer of NDV as hemagglutination units (HAU) that reduce 50% of cell population as compared to the untreated control was determined after 72 hours. TheIC50 values for cytolytic effects of NDV strain AF2240 on DBTRG.05MG cell line were 955 HAU/ml and 460 HAU/ml for the monolayer and co-culture methods, respectively. For U- 87MG cell line, the IC50 values were 380 HAU/ml and 52 HAU\ml for monolayer and co-culture methods, respectively. No significant cytolytic effect was observed on normal HCN-2 and 3T3 cell lines at the same titre used in the brain tumor cell lines. The cell proliferation rate of treated brain tumor cell lines was reduced significantly with time and titration of the virus as compared to the untreated control. It was confirmed that the mode of cell death in response to infection by NDV strain AF2240 on brain tumor cell lines was by apoptosis. Morphological features of apoptosis were observed by Phase Contrast Microscopy, Fluorescence Microscopy (Acridine Orange (AO) and Propidium Iodide (PI) staining) and Transmission Electron Microscopy. Features observed included chromatin condensation and margination along the inner nuclear membrane, cytoplasmic condensation, and membrane blebbing without disintegration of the cellular membrane. These were further confirmed with DNA laddering in agarose gel electrophoresis assay and terminal deoxyribonucleotide transferase-mediated dUTP-X nick end-labeling staining (TUNEL) assay. However, analysis of the cellular DNA content using PI showed that the virus caused an increase in sub-G1 region. The apoptosis peaks (sub-G1) found in DBTRG.05MG cells treated with NDV strain AF2240 were 18.40 and 37.40% for 24 and 48 hours, respectively whereas in U-87MG cells treated with NDV strain AF2240 the peaks were 10.29 and 19.45% for 24 and 48 hours, respectively. Early apoptosis was also observed by annexin V flow cytometry method. The amounts of apoptotic cells were 3.7 and 4.26% for DBTRG.05MG cells and U-87MG cells 6 hours post-inoculation, respectively. It was concluded that NDV strain AF2240 is a potent antitumor agent and the mode of cell death induced by this virus is apoptosis

Exploring the Excluded Stomach: A Case Series of Novel Endoscopic Techniques to Diagnose Gastric Cancer in the Excluded Stomach After Roux-en-Y Gastric Bypass Surgery

Gastric cancer is the fifth most common malignancy worldwide and the fourth leading cause of cancer-related deaths. The diagnosis is usually made by direct visualization with supporting histopathology. However, patients with gastric bypass surgery pose a challenge in diagnosis due to the difficulty in the evaluation of the excluded stomach. We present two cases of gastric cancer in the excluded stomach after Roux-en-Y gastric bypass (RYGB) surgery was diagnosed using two different endoscopic approaches

Evaluation of ultra-microscopic changes and proliferation of apoptotic glioblastoma multiforme cells induced by velogenic strain of Newcastle disease virus AF2240

Aim: Newcastle disease virus (NDV) is a member of genus Avulavirus within the family Paramyxoviridae. Interest of using NDV as an anticancer agent has arisen from its ability to kill tumor cells with limited toxicity to normal cells. Methods: In this investigation, the proliferation of brain tumor cell line, glioblastoma multiform (DBTRG.05MG) induced by NDV strain AF2240 was evaluated in-vitro, by using MTT proliferation assay. Furthermore, Cytological observations were studied using fluorescence microscopy and transmission electron microscopy, DNA laddering in agarose gel electrophoresis assay used to detect the mode of cell death and analysis of the cellular DNA content by flowcytometery. Results: MTT proliferation assay, Cytological observations using fluorescence microscopy and transmission electron microscopy show the anti-proliferation effect and apoptogenic features of NDV on DBTRG.05MG. Furthermore, analysis of the cellular DNA content showed that there was a loss of treated cells in all cell cycle phases (G1, S and G2/M) accompanied with increasing in sub-G1 region (apoptosis peak). Conclusion: It could be concluded that NDV strain AF2240 is a potent antitumor agent that induce apoptosis and its cytotoxicity increasing while increasing of time and virus titer

Antitumor Activity of Ficus deltoidea Extract on Oral Cancer: An In Vivo Study

Background. The aim of this study is to evaluate the chemopreventive and chemotherapeutic activities of Ficus deltoidea (FD) in an animal model induced for oral cancer using 4-nitroquinoline-1-oxide (4NQO). Methods. Male Sprague-Dawley (SD) rats were randomized into six groups (n = 7 per group): Group 1 (untreated group); Group 2 (control cancer group) received 4NQO only for 8 weeks in their drinking water; Groups 3 and 4 (chemopreventive) received 4NQO for 8 weeks and were simultaneously treated with FD extract at 250 and 500 mg/kg, respectively, by oral gavage; Groups 5 and 6 (chemotherapeutic) received 4NQO for 8 weeks followed by the administration of FD extract at 250 and 500 mg/kg, respectively, for another 10 weeks. The incidence of oral cancer was microscopically evaluated. Moreover, immunohistochemical expression was analysed in tongue specimens using an image analyser computer system, while the RT2 profiler PCR array method was employed for gene expression analysis. Results. The results of the present study showed a beneficial regression effect of the FD extract on tumor progression. The FD extract significantly reduced the incidence of oral squamous cell carcinoma (OSCC) from 100% to 14.3% in the high-dose groups. The immunohistochemical analysis showed that the FD extract had significantly decreased the expression of the key tumor marker cyclin D1 and had significantly increased the expression of the β-catenin and e-cadherin antibodies that are associated with enhanced cellular adhesion. Based on the gene expression analysis, FD extract had reduced the expression of the TWIST1 and RAC1 genes associated with epithelial-mesenchymal transition (EMT) and had significantly downregulated the COX-2 and EGFR genes associated with cancer angiogenesis, metastasis, and chemoresistance. Our data suggest that the FD extract exerts chemopreventive and chemotherapeutic activities in an animal model induced for oral cancer using 4NQO, thus having the potential to be developed as chemopreventive and chemotherapeutic agents

Histological, Biochemical, and Hematological Effects of Goniothalamin on Selective Internal Organs of Male Sprague-Dawley Rats

Goniothalamin (GTN) is an isolated compound from several plants of the genus Goniothalamus, and its anticancer effect against several cancers was reported. However, there is no scientific data about effects of its higher doses on internal organs. Accordingly, this study aimed to evaluate the acute and subacute effects of higher doses of GTN on the hematology, biochemistry, and histology of selected internal organs of male Sprague-Dawley rats. In acute study, 35 rats were distributed in 5 groups (n=7) which were intraperitoneally (IP) injected with a single dose of either 100, 200, 300, 400, or 500 mg/kg of GTN, while extra 7 rats serve as a normal control. In subacute study, 7 rats were IP-injected with a daily dose of 42 mg/kg of GTN for 14 days, while another 7 rats serve as a normal control group. The normal controls in both studies were IP-injected simultaneously with 2 ml/kg of 10% DMSO in PBS. At the end of both tests, rats were sacrificed to collect blood for hematology and biochemistry and harvest livers, kidneys, lungs, hearts, spleens, and brains for histology. During acute and subacute exposure, no abnormal changes were observed in the hematology, biochemistry, and histology of the internal organs. However, the 300, 400, and 500 mg/kg of GTN during acute exposure were associated with morbidities and mortalities. Ultimately, GTN could be safe up to the dose of 200 mg/kg, and the dose of 42 mg/kg of GTN was tolerated well