Mirna Profile Of Cancer Essay - 1062 Words

1.12 Aim of the study A very promising approach for understanding the involvement of a certain miRNA in the process of a disease such as cancer, is to assess the miRNA expression profile of cancer patients, and to compare with their normal counterpart is a very promising approach for understanding the involvement of a certain miRNA in a disease of interest. This may lead to the discovery of a novel prognostic or diagnostic biomarker. Although, it is believed that miRNA profile from extracted total cellular RNA is an actual indicator of RISC complex-associated miRNA, due to quick degradation of non-guide miRNA, this hypothesis was challenged by the Cullen group, when they compared the miRNA profile of the total cellular miRNAs with the†¦show more content†¦U2932 EBV positive clone A: a U2932, infected with EBV-GFP and not expressing EBNA2 [92]. SUDHL5: a B-cell line, derived from a lymph node of a 17-year-old woman with B-cell non-Hodgkin lymphoma (B-NHL) and has characteristic of GCB-DLBCL. SUDH5 EBV positive: SUDH5 cell line, infected with EBV-GFP, obtained from University of Rome. 1.12.2 RNA related techniques 1.12.2.1 RNA isolation Suspension cells (5 to 10 ml) were harvested by centrifugation at 500 x g for 5 minutes at room temperature (RT). 700 ÃŽ ¼l of Qiazol (Qiagen) was added to the pellet and incubated for 5 min at RT. Afterwards, 125 ÃŽ ¼l of chloroform was added to the sample and vortexed for 15 sec, and subsequently was again incubated for 10 min at RT. Then, the sample was centrifuged at 12000 xg for 15 min at 4 â„Æ' and the upper phase was transferred to a new 1.5 ml microcentrifuge tubes and 500 ÃŽ ¼l of isopropanol was added to the tube and incubated on ice for 15 min, followed by centrifugation at 12000 xg for 30 min in 4 â„Æ' . After that, the pellet was air dried at RT for 15 min and dissolved in 50 ÃŽ ¼l of RNase free H2O and RNA was dissolved by incubation at 55 â„Æ' for 15 min. RNA isolation for sequencing, qRT –PCR and Ago2-IP for NB was performed, using miRNeasy Mini kit, from Qiagen, according to manufacturer’s instructions, as described in the handbook forShow MoreRelatedThe Sequence Of Target Messenger Rna1604 Words   |  7 PagesMicroRNAs (miRNA) are small noncoding RNA, usually 17-25 nucleotides long that are able to bind complementary sequences of target messenger RNA (mRNA) and to induce both their degradation and translational repression (Fortunato, et al 2014). They are one of the most significant classes of non-coding RNA molecules (eg. small interfering RNA (siRNA) and ribozymes) that act within the cell. MiRNAs are also evolutionary conserved in different species from plants to humans and are encoded by their respectiveRead MoreBiology : Cell Biology, Normal Function1358 Words   |  6 PagesPM331 Essay: Exosomes, their cell biology, normal function in the cell, and role in disease. Study of exosomes has become popular in recent years, exosomes which are mainly found in body fluids such as blood and urine may play a role in cell biology and in the immune system. In eukaryotes, some cells are responsible for receiving signals via chemokines or cytokines, taking up nutrients and secreting proteins into extracellular space, endocytosis and exocytosis are then involved. Exosomes Biology: