Human Genetics And Molecular Medicine - Master Dissertation
Permanent URI for this collectionhttps://kr.cup.edu.in/handle/32116/104
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Item Potential Mitochondrial-Specific Function Of piRNAs(Central University of Punjab, 2018) Paul, Shouvik; Singh, SandeepPiwi-interacting RNAs (piRNAs) are (26-31 nt) small noncoding RNAs processed from their longer precursor transcripts with the help of Piwi proteins. There are more than 30,000 piRNA genes present in the human genome which now turns out to be emerging player in both homeostasis and diseases. Localization of piRNA and PIWI in the repeat region of the mammalian nuclear genome in germ cells has been reported, although localization and potential functional role of piRNA in the mammalian mitochondrial genome are largely unknown. We have taken 111 piRNA sequences found in the MCF-7 mitochondrial genome, which is obtained by NGS analysis for alignment study. Resulting piRNA have been aligned with DQ112870 North American Homo sapiens mitochondrion genome for studying post- transcriptional roles of piRNA.Item Generation of Rho-0 Cells using MDA-MB-231 Cell Line and Measurement of Drug Cytotoxicity(Central University of Punjab, 2018) Sharma, Bharti; Singh,SandeepThe ATP generation via Oxidative phosphorylation (OXPHOS) system located in the inner membrane of mitochondria, is regulated by the coordinated interaction between nucleus and mitochondria. In the same context, mitochondrial-depleted cell (Rho-0) can be a helpful approach to study the mitochondrial metabolism, mitochondrial role in various cellular processes such as apoptosis, mitochondrial role in various mitochondrial related disorders and cancer. To generate Rho-0 cells, EtBr mediated mtDNA depletion was done and verified by agarose gel electrophoresis. % cell viability, mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) production was measured after 24 hr treatment with 3 drugs, ?-amanitin, Doxorubicin and DCA in both parental MDA-MB-231 and Rho-0 cells. Reduced cell death and ROS production was observed in Rho-0 cells indicating the resistance against apoptosis in Rho-0 cells and demonstrating the possible role of mitochondria in intrinsic pathway of apoptosis. MMP was observed to be maintained in Rho-0 cells indicating the role of nuclear genome in the maintenance of MMP.Item Study the effect of phytochemicals phenethyl isothiocyanate (PEITC) and Quercetin on mitochondrial biogenesis in cancer and normal cell lines(Central University of Punjab, 2018) Thakur, Anchal; Chander, HarishPhytochemicals are plant-derived chemicals generally are biologically active compounds and mediate positive health benefits by targeting genes or metabolic pathways of a cell. The phytochemicals examined can be classified into main categories, such as carotenoids and polyphenols, which include phenolic acids, flavonoids and stilbenes / lignans. Quercetin is isolated from the Tridax procumbens (Linn.). Its anti-cancer activity has been well documented in vitro and in vivo. It could be pro-apoptotic as well as anti-apoptotic depending upon its concentration of it and time of exposure. Isothiocyanates are cruciferous derived phytochemicals. PEITC majorly isolated from Nasturtium officinale (watercress) has shown to mediate its anti-cancer activity through ROS-mediated pathway. It is a basic leucine zipper protein involved in protection against oxidative damage triggered by stress like injury or inflammation through regulation of the expression of anti- oxidant proteins. Under oxidative stress, inactivation of Kelch-like ECH-associated protein 1 (Keap1) occurs which is a cytosolic repressor protein that binds to Nrf2. This results in Nrf2-Keap1 complex dissociation, and hence, promoting the translocation of Nrf2 to the v nucleus where it binds to ARE (anti-oxidant response element), and induce the transcription of anti-oxidative proteins. Quercetin and PEITC treatment to the cancer cells led to decreased mitochondrial biogenesis as the NRF-2 levels diminishes as the concentration of the drug increases. The anti-oxidant levels are getting down in the cancer cells leading to ROS accumulation in the cancer cells leading ultimately to the death. Quercetin and PEITC treatment to the normal HBL-100 cells induced the mitochondrial biogenesis by increasing NRF-2 levels as the concentration of the drug increases. Confocal microscopy results also proved that treatment of quercetin or PEITC or the combination of both drugs was found to be effective in cancer cells as the mitochondria size and shape got decreased interpreted through the intensity of green dye. To conclude our study, it has been shown that quercetin and PEITC lead to increased mitochondrial biogenesis in normal cells whereas decreased mitochondrial biogenesis in cancer cells.