الفهرس 
INTRODUCTION AND AIM OF THE WORKOnly 14 pages are availabe for public view
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Abstract
Cancer is a term used for disease in which abnormal cells divide without control and are able to invade other tissues. Cancer cells can spread to other parts of the body through the blood and lymph systems. There are more than 100 different types of cancer. Most cancers are named for the organ or type of cell in which they start for example, cancer that begins in the lung is called lung cancer; cancer that begins in basal cells of the skin is called basal cell carcinoma.Paclitaxel (Taxol) is the most effective antitumor agent developed in the past three decades. It has been used for effective treatment of a variety of cancers including refractory ovarian cancer, breast cancer, non-small cell lung cancer, AIDS related Kaposi’s sarcoma, head and neck carcinoma and other cancers. Taxol inhibits cell proliferation by promoting the stabilization of microtubules at the G2-M phase of the cell cycle, by which depolymerization of microtubules to soluble tubulin is blocked. Paclitaxel stabilizes microtubules by suppressing dynamic changes leading to mitotic arrest, resulting in the failure of chromosomes to segregate, without affecting DNA, RNA or protein synthesis. Paclitaxel at low concentrations (10nM for 20 h) induces a90% mitotic block at the metaphase/anaphase transition in HeLa cells, apparently by suppressing dynamics of spindle microtubules.
Fungal endophytes are the versatile microorganisms which are widely known for the production of different metabolites. The organisms can be isolated from any part of the plant viz. leaf, root, bark, etc. from different kinds of plants. They are well recognized for the secretion
of different kinds of secondary metabolites like alkaloids, terpeniods, phenols, etc. Palm branche and strawberry fruits were employed for fungal endophyte of Alternaria sp and Cladosporium sp. were isolated for the production of Taxol.
In addition, the quantitative studies using UV suggested the presence of Taxol compared to the authentic Taxol. The compound present in all the species of Alternaria and Cladosporium confirm the presence of alkyl group attached to the aromatic ring with the presence of unsaturation groups like alkene/ alkyne side chain containing polar groups. HPLC analyses showed the concentration of Alternaria sp. was showed the less concentration of Taxol compared to the Cladosporium species.
The higher concentration of taxol extracted from samples were then exposed against A549. The in vitro anti proliferative activity studies were proved that cancer cell lines were inhibited significantly with the increasing of drug concentration. In A549 cell line more cytotoxicity effect was observed in 24hrs treatment, it also revealed that increased concentration of drug shown good toxicity over the cancer cell. This effect is due to the drug having more specific activity to cancer cell line. It was recorded that IC50 of sample was 60 mg/ml.
The novel research findings in reporting the chromatographic Analysis and Molecular studies in all species of fungi. The higher concentration of the taxol producing fungus is used for Anti-cancer activity marks the study to be worthful. Based on the conserved region of its gene, the specific primer was designed and synthesized for the amplification of the core of DNA fragment from Alternaria sp and Cladosporium sp. Following PCR amplification, approximately 650-bp products were obtained and can further be employed for genetic engineering. We conducted a qRT-PCR analysis to investigate the expression of miR-155 in A549/Taxol and A549 cell lines. As shown in (Figure 14), and the expression of miR-155 in both taxol
produced by endophytic fungi comparing with control (Figure 15). miR-
155 expression was significantly suppressed in A549/Taxol cells in
comparison to A549 cells (P<0.05), which suggested that the downregulation of miR-155 might serve an important role in suppressing the sensitivity of NSCLC cells to treatment with paclitaxel.
The proven results for their anti-cancerous role can be inferred in detailed studies like in vivo assays in future. Altogether, the study reveals novel method for the identification of Taxol produced from endophytic fungi species. Henceforth, the chosen Fungus possesses several secondary metabolites which can find application in industrial and therapeutic arenas as well as compounds for drug targets.