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आयतन 6, मुद्दा 6 (2014)

शोध आलेख

Anti-Proliferative and Apoptotic Efficacies of Ulvan Polysaccharides against Different Types of Carcinoma Cells In Vitro and In Vivo

Osama M Ahmed and Rasha R. Ahmed

Objective: This study was designed to assess the anti-proliferative and apoptotic effects of ulvan polysaccharides in vivo using Ehrlich ascites carcinoma (EAC)-bearing mice model and in vitro using hepatocarcinoma cell lines (HepG2) and colon carcinoma cell lines (HCT116).

Methods: In vivo, ulvan polysaccharide was orally administered to EAC-bearing mice at dose level of 100 mg/ kg b. w. for 2 weeks beginning from the 1st day of EAC-intraperitoneal transplantation and compared with a control given a vehicle. The expressed anti-apoptotic protein Bcl2, proapoptotic mediator p53 and DNA fragmentation marker TdT were detected by TUNEL assay. Plasma and ascites total sialic acid was determined. In vitro, the anti-tumor effect of ulvan polysaccharide against HepG2 and HCT116 was tested at 0, 12.5, 25, 50 and 100 μg/ml and IC50 was determined.

Results: The data revealed that EAC-aliquot volume, EAC-total and alive cell numbers were potentially decreased while dead cell number and percent were profoundly increased as a result of treatment with ulvan polysaccharide. The EAC-cells exhibited phenotypic signs of apoptosis after treatment with ulvan polysaccharide. The expression of proapoptotic and cell cycle arrest protein p53 in cytoplasm and nuclei and the amount of TdT in the nuclei of EAC-cells in mice treated with ulvan polysaccharide were remarkably increased while the antiapoptotic protein Bcl-2 expression was decreased. The treatment of EAC-bearing mice with ulvan polysaccharides successfully decreased plasma and ascites total sialic acid level. In vitro, the ulvan polysaccharide induced potential anti-proliferative and anti-tumor cytotoxic effects against EAC-cells, hepatoma (HepG2) and colon carcinoma (HCT116) human cell line.

Conclusion: Taken together, this study may provide evidence for the anti-tumor effects of ulvan polysaccharides which may be mediated by induction of apoptosis and suppression of cell division.

शोध आलेख

Evaluation of Anti-inflammatory and Anti-proliferative Activity of Abutilon indicum L. Plant Ethanolic Leaf Extract on Lung Cancer Cell Line A549 for System Network Studies

Kaladhar DSVGK, Swathi Saranya K, Varahalarao Vadlapudi and Nagendra Sastry Yarla

Pharmaceutically important components in medicinal plant kingdom are useful to control and cure many diseases like lung cancer. In the present study, an evaluation was performed to know the anti-inflammatory and anti-proliferative activity of ethanolic leaf extract of Abutilon indicum for potential chemopreventive agent against lung cancer. Experimentation is also conducted on lung cancer cell line A549 along with interaction studies of Apaf- 1 gene. The ethanolic leaf extract of A. indicum is showing good anti-inflammatory activity (IC50: 8.89 μg/mL) based on 5-Lipoxygenase (5-LOX) inhibition assay. The standard compound, Curcumin has shown IC50 as 8.14 μg/mL based on study. The ethanolic leaf extract of A.indicum has also shown good response on human Caucasian lung carcinoma of A549 cell line (IC50: 85.2 μg/mL) shows anti-proliferative activity. Further analysis has also shown the interaction of proteins that are involved in inflammatory and cancer response. Apoptosis-activating factor, Apaf- 1 gene increases the sensitivity of A549 cell line through interaction with proteins like CASP9, CASP3, CYCS, BCL2L1, TP53, BCL2, CASP8, HSPA4, DIABLO and CASP7. The experimental work concludes that bioactive components present in the ethanolic leaf extract of A.indicum shows good anti-inflammatory and anti-proliferative activity by inducing Apaf-1 through CASP9, CASP3, CYCS, BCL2L1, TP53, BCL2, CASP8, HSPA4, DIABLO and CASP7 network.

शोध आलेख

Improved Fat Clearance Techniques for the Examination of Breast Cancer Lymph Nodes

Fabiana Resende Rodrigues, Andréa Rodrigues Cordovil Pires, Eliene Carvalho de Fonseca, Samantha Cunha Gomes Antunes, Calina Maria Loures de Oliveira Teixeira, Licínio Esmeraldo da Silva, Nathalie Henriques Silva Canedo and Mayra Carrijo Rochael

Introduction: The isolation of lymph nodes less than 4.0 mm in diameter from the fatty tissue of surgical breast cancer specimens through the traditional method of tactile perception is difficult, time consuming and failed the identification of smaller lymph nodes. The reported numbers of lymph nodes detected through this technique vary widely in the literature demonstrating its inappropriateness. The use of fat clearance techniques however may contribute to the isolation of additional nodes, allowing for appropriate pathological staging of the disease.

Objective: To evaluate the impact of the use of fat clearance machinery and solutions in the dissection and identification of lymph nodes from axillary dissections of patients with breast cancer.

Materials and methods: Fat clearance techniques (cooker, electric spiral, modified Koren clearing solution and formaldehyde-acetic alcohol solution) were applied to 100 breast cancer axillary dissections from males and females with no age restriction over a period of two years (2009-2011).

Results: Through the use of fat clearance methods, additional 174 metastases in 564 lymph nodes were found, more than the classical method, from which 449 metastases in 1426 lymph nodes were identified. There was no statistical difference between the fat clearance methods; however the cooker method demonstrated greater efficiency than the modified Koren clearing solution method. Conclusion: The fat clearance methods described here have various advantages over the classical method. These methods are viable, rapid, practical, inexpensive and do not alter the quality of histological analysis or immunohistochemical reactions. The findings show that examination of a minimum of 20 isolated lymph nodes is necessary for optimal identification and diagnosis of lymph node metastases. The main outcome from this study were the immediate benefits generated by adopting new technical procedures resulting in a change in pathological staging classification in 14% of the patients.

शोध आलेख

A Comprehensive Approach to Patient-individual Glioblastoma Multiforme Model Establishment

Christina Susanne Mullins, Björn Schneider, Anne Lehmann, Florian Stockhammer, Sascha Mann, Carl-Friedrich Classen and Michael Linnebacher

Patient-individual tumor models for Glioblastoma Multiforme (GBM) are important not only for basic and translational research but also for the development and improvement of optimal and individualized treatment strategies. The model that has gained widest acceptance is the primary cell culture model. The laborious and time consuming process is rewarded with a relative high initial success rate (about 60%). We here describe and evaluate an extended biobanking methodology to simplify sample collection and model establishment. GBM resection specimen were collected ad hoc, partially prepared fresh for modeling, snap frozen for molecular testing and frozen down vitally.

The established models were subject to subsequent detailed characterization in direct comparison to the patients´ tumors. Generally, molecular characteristics such as mutations, gene amplifications and epigenetic alterations were maintained in the models. Immortality, neuronal origin and stem cell characteristics of the cell lines could be demonstrated. Extensive drug sensitivity screens were performed. These well-defined patient-individual models are ideal for establishment of individualized therapy approaches and enable testing of immunological strategies.

Our extended biobanking procedure facilitates collection, long-time storage and propagation (modeling) of clinical GBM specimens (potentially also from distant centers) for basic research, (pre-) clinical studying of novel therapies and individual response prediction.

समीक्षा लेख

Gene Silencing – Targeting at Gene Methylation and Demethylation Patterns for the Arrest of Cancer Progression

Rekha Govindan, Sourabh Suran and Sanjeeva Metikala

The sudden occurrence or silent recurrence of tumor has been found to result from long term molecular changes at DNA level, that eventually lead to the abnormal expansion of cancer cells. One such molecular change is accounted by a distinct promoter DNA hyper-methylation pattern in certain tumor suppressing genes that are usually the check points for the regulation of gene expression in normal and progenitor cells. The hyper methylated promoter regions may account for the transcriptional inactiveness of the tumor suppressor genes. This possibility, coupled with the reversible nature of epigenetics such as demethylation has enormous significance for the prevention and control of cancer. In such circumstances, the use of demethylating drugs may re-express those genes that were silenced by aberrant DNA methylation. Understanding these epigenetic changes may help in developing specific markers to identify the molecular pathways of tumorigenesis and for deriving sensitive molecular detection strategies for a variety of tumor type.

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