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Clinical Applications of siRNA
Published in Yashwant V. Pathak, Gene Delivery Systems, 2022
Seth Kwabena Amponsah, Ismaila Adams, Kwasi Agyei Bugyei
EGFRs are good targets for inhibiting tumor angiogenesis. Some carcinogenic EGFR genes have been coupled with siRNA in lung cancer models, and this has led to significant therapeutic impact (17). Additionally, the use of siRNA to target p53 for the treatment of lung cancers has recently received a lot of attention (18). Lung cancers can be treated with a combination of miR-34a (which restores p53 downstream) and siRNA (19). This combination has the tendency to increase apoptosis in cells and decrease tumor volume (19). HDAC regulates protein expression by interacting with cancer-related genes and preventing tumor development. In an orthotropic xenograft model, HDAC siRNA/lipid nanoparticles were found to be beneficial in decreasing the development of liver tumors (20). Furthermore, Itgb1 is important for hepatoma cell proliferation. Experiments have demonstrated that siRNAs targeting Itgb1 can slow the progression of hepatocellular carcinoma (21–22). In hepatocellular and breast cancer cells, human telomerase reverse transcriptase (hTERT) is abundantly expressed and active (23). Telomerase activity can be reduced by using hTERT-siRNA that targets bioreducible polyethylenimine (SS-PEI) (24). It is therefore not surprising that about one-third of siRNA-based medicinal interventions target cancers (25).
Application of Carbon Nanotubes in Cancer Vaccines as Drug Delivery Tools
Published in Loutfy H. Madkour, Nanoparticle-Based Drug Delivery in Cancer Treatment, 2022
The way the CNTs interact with nucleic acids has been extensively studied for their potential applications. Both antisense oligonucleotides and small interfering RNA (siRNA) are very promising fragments for gene silencing, applicable for the treatment of many diseases. They can in fact inhibit protein expression, potentially blocking many cellular pathways. Cancer therapy is one of the possible applications, when the targets are oncogenes or genes involved, for instance, in angiogenesis or chemotherapy resistance. One of the first studies in this field was performed a few years ago [181]. Cationic SWCNTs were used to complex siRNA, able to silence the expression of telomerase reverse transcriptase and thus inhibit cell growth. This activity was proved in vitro, on different cell lines, both murine and human, and in vivo, after intra-tumor injection in xenografted mice.
Carbon Nanotubes For Biomedical Applications
Published in Shrikaant Kulkarni, Iuliana Stoica, A.K. Haghi, Carbon Nanotubes for a Green Environment, 2022
An approach first of its kind for genetic material delivery was undertaken by Lu et al. who chelated RNA polymer poly(rU) with SWCNTs with the help of interactions nonspecific in nature.33 The translocation across MCF7 breast cancer cells was found to take place and the genetic material was embedded across the membranes both cellular and nuclear. Derivatives of SWCNTs, synthesized using amidation of the CNTs carboxylic groups with a chain possessing an ammonium group at the terminal position, were examined as telomerase reverse transcriptase RNA (TERT-siRNA) carrier into cells.34 The performance was evaluated considering the spreading of tumor cells (Lewis lung cancer and HeLa cells), in vitro and in vivo. the targeted gene is silenced shows an improved intracellular delivery of siRNA, which gets released at the site from CNTs and can cause the necessary effect.
Serum from differently exercised subjects induces myogenic differentiation in LHCN-M2 human myoblasts
Published in Journal of Sports Sciences, 2018
D. Vitucci, E. Imperlini, R. Arcone, A. Alfieri, A. Canciello, L. Russomando, D. Martone, A. Cola, G. Labruna, S. Orrù, D. Tafuri, A. Mancini, P. Buono
The human myoblast cell line, LHCN-M2, is the first species-specific system of immortalized cells able to overcome cellular senescence through the expression of the human telomerase reverse transcriptase and cyclin-dependent kinase 4 (Zhu et al., 2007). Here, we demonstrate that when LHCN-M2 cells are induced to differentiate, they show morphological modifications, namely, an increase in the fusion index and in the expression of the muscle-specific differentiation markers, CK activity, myogenin and MyHC-β (Diel, Baadners, Schlupmann, Velders, & Schwarz, 2008; Shen et al., 2003). We also found that p62 and Bcl-2, which are protein markers of the autophagy and apoptosis pathways respectively, were abundantly expressed in LHCN-M2 cells during the early stages of differentiation, and decreased in the late stages of differentiation, similar to what occurs in murine myoblasts (Lindqvist & Vaux, 2014; McMillan & Quadrilatero, 2014; Schoneich, Dremina, Galeva, & Sharov, 2014; Sin et al., 2016). Furthermore, apoptosis is regulated by caspases, which when activated, induce cleavage of the PARP-1 protein (Anderson, Russell, & Foletta, 2015; Olah et al., 2015). We found no change in PARP-1 expression in LHCN-M2 cells 4 days after differentiation induction. Thus, our results indicate that the LHCN-M2 cell line is able to differentiate in mature myotubes when cultured in DM for 2, 4 and 7 days, and to express early (myotube formation, creatine kinase activity and myogenin) and late (MyHC-β) muscle-specific markers.
Efficient expression of recombinant human telomerase inhibitor 1 (hPinX1) in Pichia pastoris
Published in Preparative Biochemistry and Biotechnology, 2018
Yagmur Unver, Melike Yildiz, Deryanur Kilic, Mesut Taskin, Abdulhadi Firat, Hakan Askin
Human telomerase adds 5′-TTAGGG-3′ repeats onto the ends of human chromosomes and is a ribonucleoprotein enzyme complex located in the core of telomere. This complex provides a telomere persistence mechanism for a grand majority (∼90%) of cancers.[1] Telomerase contains several subunits. Human telomerase reverse transcriptase (hTERT) is its catalytic subunit and contains four C-terminal and two conserved N-terminal domains responsible for telomerase catalytic activity. hTERC subunit attends as a RNA template while telomere elongation. For telomerase activity, hTERT expression level may be a simulated marker. This foresight is widely accepted.[2,3]
Toward an integrated framework for assessing micropollutants in marine mammals: Challenges, progress, and opportunities
Published in Critical Reviews in Environmental Science and Technology, 2021
Edmond Sanganyado, Ran Bi, Charles Teta, Lucas Buruaem Moreira, Xiaoxuan Yu, Sun Yajing, Tatenda Dalu, Imran Rashid Rajput, Wenhua Liu
The utility of marine mammal cell cultures in effect assessment is often affected by their lifespan, the physicochemical properties of test chemicals, the fact that not all cells are culturable, and the limitations of extrapolating cell models to whole animals. In addition, some mammalian cells cannot not proliferate under cell culture conditions. Culturable marine mammal cells often proliferate for around 40–60 cycles before they reach senescence. The cell cycle and senescence can be extended by introducing mutations and chromosomal abnormalities or mutations that permit indefinite proliferation. Previous studies transfected simian virus large T tumor antigen (SV40T) to extend the lifespan of Atlantic bottlenose dolphin (Yu et al., 2005), pygmy killer whale (Yajing et al., 2018), Indo-Pacific hump back dolphin fibroblast cells (Jin et al., 2013). Interestingly, the lifespan of Indo-Pacific humpback dolphin cell culture could not be extended using SV40T; however, it was succesfully extended by blocking telomere shortening using telomerase reverse transcriptase (Burkard et al., 2019). Furthermore, depending on their physicochemical properties, micropollutants can be lost through volatilization, adsorption, photolysis, and hydrolysis resulting in different toxicity values (Schreiber et al., 2008; Tanneberger et al., 2010). Passive dosing using silicone O-rings has been shown to curb the loss and heterogenous distribution of test chemical during effect assessment of hexachlorobenzene to humpback whale fibroblast cell line (Maner et al., 2019). Furthermore, cell cultures often overlook the effects of multiple organs on the adsorption, distribution, metabolism, and elimination of the micropollutants. Hence, despite offering critical mechanistic data on the toxicity of micropollutants, additional studies are required on extending lifespan and improving the accuracy of cell lines in effect assessment.