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Methods in Molecular Biology (Clifton,... 2018RNA interference (RNAi) is a biological process by which double-stranded RNA (dsRNA) induces sequence-specific gene silencing by targeting mRNA for degradation. As a... (Review)
Review
RNA interference (RNAi) is a biological process by which double-stranded RNA (dsRNA) induces sequence-specific gene silencing by targeting mRNA for degradation. As a tool for knocking down the expression of individual genes posttranscriptionally, RNAi has been widely used to study the cellular function of genes. In this chapter, I describe procedures for using gene-specific, synthetic, short interfering RNA (siRNA) to induce gene silencing in mammalian cells. Protocols for using lipid-based transfection reagents and electroporation techniques are provided. Potential challenges and problems associated with the siRNA technology are also discussed.
Topics: Animals; Electroporation; Gene Knockdown Techniques; Humans; RNA Interference; RNA Stability; Transfection
PubMed: 29423805
DOI: 10.1007/978-1-4939-7471-9_16 -
Diagnostic and Interventional Imaging Nov 2022Thermal tumor ablation techniques including radiofrequency, microwave, LASER, high-intensity focused ultrasound and cryoablation are routinely used to treated liver,... (Review)
Review
Thermal tumor ablation techniques including radiofrequency, microwave, LASER, high-intensity focused ultrasound and cryoablation are routinely used to treated liver, kidney, bone, or lung tumors. However, all these techniques are thermal and can therefore be affected by heat sink effect, which can lead to incomplete ablation, and thermal injuries of non-targeted tissues are possible. Under certain conditions, high voltage pulsed electric field can induce formation of pores in the cell membrane. This phenomenon, called electropermeabilization, is also known as "electroporation". Under certain conditions, electroporation can be irreversible, leading to cell death. Irreversible electroporation has demonstrated efficacy for the treatment of liver and prostate cancers, whereas data are scarce regarding pancreatic and renal cancers. During reversible electroporation, transient cell permeability can be used to introduce cytotoxic drugs into tumor cells (commonly bleomycin or cisplatin). Reversible electroporation used in conjunction with cytotoxic drugs shows promise in terms of oncological response, particularly for solid cutaneous and subcutaneous tumors such as melanoma. Irreversible and reversible electroporation are both not thermal ablation techniques and therefore open a new promising horizon for tumor ablation.
Topics: Humans; Electrochemotherapy; Medical Oncology; Electroporation; Neoplasms; Antineoplastic Agents
PubMed: 36266192
DOI: 10.1016/j.diii.2022.09.009 -
Bioelectrochemistry (Amsterdam,... Oct 2021Exposure of cells to high voltage electric pulses increases transiently membrane permeability through membrane electroporation. Electroporation can be reversible and is... (Review)
Review
Exposure of cells to high voltage electric pulses increases transiently membrane permeability through membrane electroporation. Electroporation can be reversible and is used in gene transfer and enhanced drug delivery but can also lead to cell death. Electroporation resulting in cell death (termed as irreversible electroporation) has been successfully used as a new non-thermal ablation method of soft tissue such as tumours or arrhythmogenic heart tissue. Even though the mechanisms of cell death can influence the outcome of electroporation-based treatments due to use of different electric pulse parameters and conditions, these are not elucidated yet. We review the mechanisms of cell death after electroporation reported in literature, cell injuries that may lead to cell death after electroporation and membrane repair mechanisms involved. The knowledge of membrane repair and cell death mechanisms after cell exposure to electric pulses, targets of electric field in cells need to be identified to optimize existing and develop of new electroporation-based techniques used in medicine, biotechnology, and food technology.
Topics: Animals; Cell Death; Electroporation
PubMed: 34147013
DOI: 10.1016/j.bioelechem.2021.107871 -
Bioelectrochemistry (Amsterdam,... Dec 2020
Topics: Animals; Biosensing Techniques; Electrochemistry; Electroporation; Energy Metabolism; Humans; Protein Engineering; Proteins
PubMed: 32645566
DOI: 10.1016/j.bioelechem.2020.107554 -
Molecules (Basel, Switzerland) Apr 2022Gynecological carcinomas affect an increasing number of women and are associated with poor prognosis. The gold standard treatment plan is mainly based on surgical... (Review)
Review
Gynecological carcinomas affect an increasing number of women and are associated with poor prognosis. The gold standard treatment plan is mainly based on surgical resection and subsequent chemotherapy with cisplatin, 5-fluorouracil, anthracyclines, or taxanes. Unfortunately, this treatment is becoming less effective and is associated with many side effects that negatively affect patients' physical and mental well-being. Electroporation based on tumor exposure to electric pulses enables reduction in cytotoxic drugs dose while increasing their effectiveness. EP-based treatment methods have received more and more interest in recent years and are the subject of a large number of scientific studies. Some of them show promising therapeutic potential without using any cytotoxic drugs or molecules already present in the human body (e.g., calcium electroporation). This literature review aims to present the fundamental mechanisms responsible for the course of EP-based therapies and the current state of knowledge in the field of their application in the treatment of gynecological neoplasms.
Topics: Antineoplastic Agents; Breast Neoplasms; Cisplatin; Electrochemotherapy; Electroporation; Female; Humans
PubMed: 35458673
DOI: 10.3390/molecules27082476 -
Journal of B.U.ON. : Official Journal... 2019Sarcomas arise from uncontrolled cell growth in tissues of mesodermal origins, such as connective tissue, bone, cartilage, fat or muscle. Surgical resection is still... (Review)
Review
Sarcomas arise from uncontrolled cell growth in tissues of mesodermal origins, such as connective tissue, bone, cartilage, fat or muscle. Surgical resection is still considered the cornerstone in the treatment of sarcomas. However, in many cases where tumor is adjacent to vital structures like major vessels, other treatment modalities may be implemented. Irreversible electroporation (IRE), a new form of ablative technique has been introduced lately in the treatment of several types of sarcomas. Irreversible electroporation has shown promising results and survival improvement in primarily inoperable solid tumors and locally advanced cancers, including prostate, kidney, liver, bone and pancreatic cancers in close proximity to important structures. The aim of this review was to sum up the current knowledge and the future perspectives of the usage of IRE in the management of sarcomas. Our study indicates that IRE could possibly represent a potential therapeutic option in patients with advanced or metastatic sarcoma, when surgery is not indicated.
Topics: Ablation Techniques; Electroporation; Humans; Male; Sarcoma; Soft Tissue Neoplasms; Treatment Outcome
PubMed: 31646777
DOI: No ID Found -
Bioelectrochemistry (Amsterdam,... Oct 2022Current-Clamp electroporation refers to the application of a constant current across a membrane which results in voltage fluctuations due to the creation of...
Current-Clamp electroporation refers to the application of a constant current across a membrane which results in voltage fluctuations due to the creation of electropores. This method allows for the measurement of electroporation across a long timescale (minutes) and facilitates the comparison between experimental and theoretical studies. Of particular interest is the claim in the literature that current-clamp electroporation results in the creation of a single pore. We simulated current-clamp electroporation using the Smoluchowski and Langevin equations and identified two possible mechanisms to explain the observed voltage fluctuations. The voltage fluctuations may be due to a single pore or a few pores growing and shrinking via a negative feedback mechanism or the opening and closing of pores in a larger population of pores. Our results suggest that current-clamp conditions do not necessarily result in the creation of a single pore. Additionally, we showed that the Langevin model is more accurate than the Smoluchowski model under conditions where there are only a few pores.
Topics: Computer Simulation; Electroporation; Models, Theoretical
PubMed: 35691267
DOI: 10.1016/j.bioelechem.2022.108162 -
The Journal of Membrane Biology Oct 2016Phylogenetic studies reveal that horizontal gene transfer (HGT) plays a prominent role in evolution and genetic variability of life. Five biotic mechanisms of HGT among... (Review)
Review
Phylogenetic studies reveal that horizontal gene transfer (HGT) plays a prominent role in evolution and genetic variability of life. Five biotic mechanisms of HGT among prokaryotic organisms have been extensively characterized: conjugation, competence, transduction, gene transfer agent particles, and transitory fusion with recombination, but it is not known whether they can account for all natural HGT. It is even less clear how HGT could have occurred before any of these mechanisms had developed. Here, we consider contemporary conditions and experiments on microorganisms to estimate possible roles of abiotic HGT-currently and throughout evolution. Candidate mechanisms include freeze-and-thaw, microbeads-agitation, and electroporation-based transformation, and we posit that these laboratory techniques have analogues in nature acting as mechanisms of abiotic HGT: freeze-and-thaw cycles in polar waters, agitation by sand at foreshores and riverbeds, and lightning-triggered electroporation in near-surface aqueous habitats. We derive conservative order-of-magnitude estimates for rates of microorganisms subjected to freeze-and-thaw cycles, sand agitation, and lightning-triggered electroporation, at 10, 10, and 10 per year, respectively. Considering the yield of viable transformants, which is by far the highest in electroporation, we argue this may still favor lightning-triggered transformation over the other two mechanisms. Electroporation-based gene transfer also appears to be the most general of these abiotic candidates, and perhaps even of all known HGT mechanisms. Future studies should provide improved estimates of gene transfer rates and cell viability, currently and in the past, but to assess the importance of abiotic HGT in nature will likely require substantial progress-also in knowledge of biotic HGT.
Topics: Electroporation; Gene Transfer, Horizontal; Transformation, Genetic
PubMed: 27067073
DOI: 10.1007/s00232-016-9897-y -
Technology in Cancer Research &... Jan 2019Irreversible electroporation has raised great interest in the past decade as a means of destroying cancers in a way that does not involve heat. Irreversible... (Review)
Review
Irreversible electroporation has raised great interest in the past decade as a means of destroying cancers in a way that does not involve heat. Irreversible electroporation is a novel ablation technology that uses short high-voltage electrical pulses to enhance the permeability of tumor cell membranes and generate irreversible nano-sized structural defects or pores, thus leading to cell death. Irreversible electroporation has many advantages over thermal therapies due to its nonthermal mechanism: (1) reduced risk of injury to surrounding organs and (2) no "heat-sink" effect due to nearby blood vessels. However, so far, it has been difficult for irreversible electroporation to completely ablate large tumors (eg, >3 cm in diameter). In order to overcome this problem, many preclinical and clinical studies have been performed to improve the efficacy of IRE in the treatment of large size of tumors through a chemical perspective. Due to the distribution of electric field, irreversible electroporation region, reversible electroporation region, and intact region can be found in the treatment of irreversible electroporation. Thus, 2 types of chemical enhancements of irreversible electroporation were discussed in the article, such as the reversible electroporation region enhanced and the irreversible electroporation region enhanced. Specifically, the state-of-the-art results regarding the following approaches that have the potential to be used in the enhancement of irreversible electroporation were systematically reviewed in the article, including (1) combination with cytotoxic drugs, (2) calcium electroporation, (3) modification of cell membrane, and (4) modification of the tumor cell microenvironment. In the end, we concluded with 4 issues that should be addressed in the future for improving irreversible electroporation further in a chemical way.
Topics: Algorithms; Animals; Antineoplastic Agents; Calcium; Cell Survival; Cellular Microenvironment; Electroporation; Humans; Models, Theoretical; Surface-Active Agents
PubMed: 31500518
DOI: 10.1177/1533033819874128 -
Bioelectrochemistry (Amsterdam,... Oct 2022Electroporation has become a powerful tool for nonviral delivery of various biomolecules such as nucleic acids, proteins, and chemotherapeutic drugs to virtually any...
Electroporation has become a powerful tool for nonviral delivery of various biomolecules such as nucleic acids, proteins, and chemotherapeutic drugs to virtually any living cell by exposing the cell membrane to an intense pulsed electric field. Different multiphysics and multiscale models have been developed to describe the phenomenon of electroporation and predict molecular transport through the electroporated membrane. In this paper, we critically examine the existing mechanistic, single-cell models which allow spatially and temporally resolved numerical simulations of electroporation-induced transmembrane transport of small molecules by confronting them with different experimental measurements. Furthermore, we assess whether any of the proposed models is universal enough to describe the associated transmembrane transport in general for all the different pulse parameters and small molecules used in electroporation applications. We show that none of the tested models can be universally applied to the full range of experimental measurements. Even more importantly, we show that none of the models has been compared to sufficient amount of experimental data to confirm the model validity. Finally, we provide guidelines and recommendations on how to design and report experiments that can be used to validate an electroporation model and how to improve the development of mechanistic models.
Topics: Biological Transport; Cell Membrane; Electricity; Electroporation; Models, Biological
PubMed: 35932533
DOI: 10.1016/j.bioelechem.2022.108216