Informatics Publishing Limited

Namrata Dwivedi ¹, Bhavna Dwivedi ¹, Skand Mishra ¹, Yogeshwer Shukla ²

Affiliations
1 Department of Botany, Govt. New Science College, Rewa (M.P)-486001, IN
2 CSIR-Indian Institute of Toxicology Research, Lucknow, IN

Abstract

Some naturally occurring chemical agents in the human diet can reduce cancer risk such as, Lupeol (Lup-20(29)-en-3-b-ol), a pentacyclic triterpene principally present in fruits such as mango, olive, strawberry, and grapes, figs, etc. have been used to treat several ailments. Lupeol has been shown to have analgesic, anti-inflammatory, antioxidant, and cytoprotective properties along with antimutagenic and hypotensive activities in in vitro and in vivo systems. Several scientists have reported that lupeol exerts antitumor effects by triggering apoptosis in cancer cells aside from affecting normal cells.The topical application of lupeol on CD-1 mouse skin showed reduced 12-O-tetradecanoyl-phorbol-13- acetate-induced tumor incidence by the inhibition of phosphatidyl inositol 3-kinase (PI3K), Akt, Nuclear factor kappa B (NFκB) and activation of Inhibitory-kB kinase-((IKK-(α).However, in vitro studies on lupeol are limited illustrating an unclear mechanism by which it imparts apoptosis. Lupeol also know to induce apoptotic death of human pancreatic adenocarcinoma cells through mitochondria mediated pathway with significant decrease of Bcl-2 and increase of Bax and caspases.In our quest to dissect the molecular mechanism of lupeol induced apoptosis and inhibition of cancer cells growth, we investigated the regulatory effects of lupeol on activation of apoptotic machinery in human lung carcinoma A549 cells. It is noteworthy that, Lupeol at its effective therapeutic doses exhibit no toxicity to normal cells and tissues. Our study focuses to determine the cancer chemotherapeutics potential of Lupeol for the treatment of lung cancer.

Keywords

Lupeol, Apoptosis, Lung Cancer, Antimutagenic.

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Bhavna Dwivedi ¹, P. K. Mishra ², S. K. Mishra ¹

Affiliations
1 Department of Biotechnology, Govt. New Science College, Rewa 486491 (MP), IN
2 Bhopal Memorial Hospital and Research Centre, Bhopal, IN

Abstract

Living cells sometimes encounter continuous or accidental exposure of genotoxic agents which misbalances the integrity of subcellular components. DNA damage by these genotoxic agents initiates various cellular responses. These cellular responses include sensors, transducers and effectors. Sensor protein act at the frontline of the DNA response mechanism. The transducer amplifies the intracellular signals to the down stream regulatory molecules. Then the effector molecules progresses towards the execution of cell cycle arrest, DNA repair or apoptosis. The most important sensor proteins are ATM and ATR. One of the most imperative substrate that is phosphorylated by these protein kinases is γ-H2AX which is a vital component for nuclear foci formation to execute DNA repair mechanisms. This complete process is under tight regulation and any imbalance in these processes may lead to hypersensitivity to cellular stress and susceptibility to DNA damage, genomic effects and resistance to apoptosis, thus initiating the cascade of carcinogenesis. There are various agents that modifies DNA molecule, for example; environmental isocyanates and their derivatives that have capability to cause toxicogenomic effects. Isocyanates are thus becoming of interest in the field of genetic toxicology as they may react with DNA to produce DNA damage.

Keywords

DNA Damage, Genotoxic Agent, ATM.

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Niharika Mishra ¹, Riyazul Hasan Khan ², Bhavna Dwivedi ¹, Skand Kumar Mishra ¹

Affiliations
1 Department of Botany, Govt. New Science College, Rewa (M.P), IN
2 Genentech Laboratories, Biotech Park, Lucknow, IN

Abstract

Glyceraldehyde-3-phosphate dehydrogenase (GPD, E.C. 1.2.1.12) is one of the key enzymes in the Embden Meyerhof Parnas or glocolysis pathway. It catalyzes phosphorylation of glyceraldehyde-3-phosphate to produce 1, 3-diphosphoglycerate. GPD is a tetrameric enzyme composed of four identical subunits. It is an essential enzyme used to maintain life activities through contributing in thisway to the formation of ATP and providing additional energy to the cell by reducing NADH to NAD+ and H+ upon its action. Moreover, GPD protein also has many other important functions, such as abiotic stress tolerance (Liu and Yang, 2005).The pUC18 vector was isolated from E.coli Top10 strain. It is a prokaryotic vector carrying a multiple cloning site with 13 unique restriction sites. This multiple cloning sites are located within the lacZ’ gene resulting in the disruption of β-galactosidase activity by cloned inserts allowing blue/white selection. The vector pUC18 was restricted using the restriction enzyme Eco.RI. Further it was thymidized to provide sticky ends so that desired gene can be inserted in it. The GAPDH gene was amplified using PCR. Then it was inserted/ligated into cloning vector pUC18.It was then transformed into the suitable competent cells which were freshly prepared using JM107 strain of E.coli which is devoid of plasmid. Inside the host cell the recombinant DNA had undergone replication; thus, a bacterial host gave rise to a colony of cells containing the cloned target gene i.e. GAPDH. Various screening methods could be used to identify such colonies, enabling them to be selected and cultured.

Keywords

β–Galactosidase, Glyceraldehyde 3-Phosphate Dehydrogenase, PCR.

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