subject:(Cell junctions Research)

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East Carolina University

1. Hoggard, John Elliott. Claudin-7 increases chemosensitivity to cisplatin in human NCI-H522 lung cancer cells.

Degree: MS, Biology, 2010, East Carolina University

► Lung cancer is a leading cause of cancer death for both men and women worldwide. The 5-year survival rate for lung cancer is only 15%,… (more)

▼ Lung cancer is a leading cause of cancer death for both men and women worldwide. The 5-year survival rate for lung cancer is only 15%, and it accounts for approximately 1.4 million deaths every year. Recently, it has been reported that changes in expression level of claudin proteins, a family of tight junction integral membrane proteins, has been associated with several types of cancer tissues. Claudin proteins are vital components of the tight junction complex, playing a crucial role in maintaining cell polarity, adhesion, and paracellular permeability. Studies have illustrated that claudin-7 expression is lost in head, neck, invasive breast cancer, and certain types of lung cancer. The objective of this study was to investigate apoptosis in a human lung adenocarcinoma cancer cell line transfected with claudin-7. We transfected claudin-7 cDNA tagged with myc into human NCI-H522 lung cancer cells, which have no detectable expression of claudin-7 protein. H522 cells transfected with vector alone were used as a control. Flow cytometry analysis demonstrated that cells transfected with claudin-7 had a significantly higher percentage of cell apoptosis when compared to that of vector transfected cell population. The cell viability by MTT, (3-(4,5-Dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide, a tetrazole), assay was significantly decreased and cell apoptosis by Annexin V assay was significantly increased in claudin-7 transfected cells compared to that of vector transfected cells after cisplatin treatment. Cisplatin is an anti-cancer drug clinically used to treat tumors in several tissues including lung tumors. Furthermore, we found that claudin-7 transfected cells exhibited upregulation of caspase-3, -8, and -9. Also, the phosphorylation of claudin-7 in the H522 cells was examined, and analyses revealed that claudin-7 was phosphorylated at serine 204 by protein kinase C. Comparison of phosphorylated and non- phosphorylated claudin-7 in the H522 cells showed decreased cell viability by the MTT assay suggesting that phosphorylation increases chemosensitivity to cisplatin treatment. We demonstrated that claudin-7 expression in H522 lung cancer cells increases chemosensitivity to cisplatin through increased activation of caspase pathways.Â Â Advisors/Committee Members: Chen, Yan-Hua (advisor).

Subjects/Keywords: Biology; Biology, Cellular; Cellular biology; Tight junctions (Cell biology) – Research; Junctional complexes (Epithelium) – Research; Membrane proteins – Research; Apoptosis – Research; Cisplatin – Research; Lungs – Cancer – Research

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Hoggard, J. E. (2010). Claudin-7 increases chemosensitivity to cisplatin in human NCI-H522 lung cancer cells. (Masters Thesis). East Carolina University. Retrieved from http://hdl.handle.net/10342/3146

Chicago Manual of Style (16^th Edition):

Hoggard, John Elliott. “Claudin-7 increases chemosensitivity to cisplatin in human NCI-H522 lung cancer cells.” 2010. Masters Thesis, East Carolina University. Accessed April 22, 2021. http://hdl.handle.net/10342/3146.

MLA Handbook (7^th Edition):

Hoggard, John Elliott. “Claudin-7 increases chemosensitivity to cisplatin in human NCI-H522 lung cancer cells.” 2010. Web. 22 Apr 2021.

Vancouver:

Hoggard JE. Claudin-7 increases chemosensitivity to cisplatin in human NCI-H522 lung cancer cells. [Internet] [Masters thesis]. East Carolina University; 2010. [cited 2021 Apr 22]. Available from: http://hdl.handle.net/10342/3146.

Council of Science Editors:

Hoggard JE. Claudin-7 increases chemosensitivity to cisplatin in human NCI-H522 lung cancer cells. [Masters Thesis]. East Carolina University; 2010. Available from: http://hdl.handle.net/10342/3146

IUPUI

2. Lewis, Shanta. Effects of carbon nanotubes on barrier epithelial cells via effects on lipid bilayers.

Degree: 2013, IUPUI

URL: http://hdl.handle.net/1805/5611

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Indiana University-Purdue University Indianapolis (IUPUI)

Carbon nanotubes (CNTs) are one of the most common nanoparticles (NP) found in workplace air. Therefore, there is a strong… (more)

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Indiana University-Purdue University Indianapolis (IUPUI)

Carbon nanotubes (CNTs) are one of the most common nanoparticles (NP) found in workplace air. Therefore, there is a strong chance that these NP will enter the human body. They have similar physical properties to asbestos, a known toxic material, yet there is limited evidence showing that CNTs may be hazardous to human barrier epithelia. In previous studies done in our laboratory, the effects of CNTs on the barrier function in the human airway epithelial cell line (Calu-3) were measured. Measurements were done using electrophysiology, a technique which measures both transepithelial electrical resistance (TEER), a measure of monolayer integrity, and short circuit current (SCC) which is a measure of vectorial ion transport across the cell monolayer. The research findings showed that select physiologically relevant concentrations of long single-wall (SW) and multi-wall (MW) CNTs significantly decreased the stimulated SCC of the Calu-3 cells compared to untreated cultures. Calu-3 cells showed decreases in TEER when incubated for 48 hours (h) with concentrations of MWCNT ranging from 4µg/cm2 to 0.4ng/cm2 and SWCNT ranging from 4µg/cm2 to 0.04ng/cm2. The impaired cellular function, despite sustained cell viability, led us to investigate the mechanism by which the CNTs were affecting the cell membrane. We investigated the interaction of short MWCNTs with model lipid membranes using an ion channel amplifier, Planar Bilayer Workstation. Membranes were synthesized using neutral diphytanoylphosphatidylcholine (DPhPC) and negatively charged diphytanoylphosphatidylserine (DPhPS) lipids. Gramicidin A (GA), an ion channel reporter protein, was used to measure changes in ion channel conductance due to CNT exposures. Synthetic membranes exposed to CNTs allowed bursts of currents to cross the membrane when they were added to the membrane buffer system. When added to the membrane in the presence of GA, they distorted channel formation and reduced membrane stability.

Advisors/Committee Members: Blazer-Yost, Bonnie, Petrache, Horia, Witzmann, F. A. (Frank A.), Atkinson, Simon.

Subjects/Keywords: Carbon nanotubes; Multi-wall carbon nanotubes; single-wall carbon nanotubes; T84; Calu-3; Black lipid membranes; Electrophysiology; Ion channels; Gramicidin A; Short circuit current; Carbon nanotubes – Research – Analysis; Organic compounds – Synthesis; Nanostructured materials – Research – Analysis; Electrophysiology – Technique; Ion channels – Research – Analysis; Short circuits; Epithelium; Gramicidins; Cyclic peptides – Synthesis; Tight junctions (Cell biology); Bilayer lipid membranes; Biological transport; Membranes (Biology)

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Lewis, S. (2013). Effects of carbon nanotubes on barrier epithelial cells via effects on lipid bilayers. (Thesis). IUPUI. Retrieved from http://hdl.handle.net/1805/5611

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Lewis, Shanta. “Effects of carbon nanotubes on barrier epithelial cells via effects on lipid bilayers.” 2013. Thesis, IUPUI. Accessed April 22, 2021. http://hdl.handle.net/1805/5611.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Lewis, Shanta. “Effects of carbon nanotubes on barrier epithelial cells via effects on lipid bilayers.” 2013. Web. 22 Apr 2021.

Vancouver:

Lewis S. Effects of carbon nanotubes on barrier epithelial cells via effects on lipid bilayers. [Internet] [Thesis]. IUPUI; 2013. [cited 2021 Apr 22]. Available from: http://hdl.handle.net/1805/5611.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Lewis S. Effects of carbon nanotubes on barrier epithelial cells via effects on lipid bilayers. [Thesis]. IUPUI; 2013. Available from: http://hdl.handle.net/1805/5611

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

3. Oruganti, Prasad. Drug Transport in Cell Preparations with Diffusional Dosing and Temporal Ratiometry.

Degree: PhD, Biomedical Engineering, 2010, Case Western Reserve University School of Graduate Studies

URL: http://rave.ohiolink.edu/etdc/view?acc_num=case1269536608

► The introduction of specific molecules into single cells is a widely used approach to probe cellular mechanisms. For example, to better understand the complex… (more)

▼ The introduction of specific molecules into single cells is a widely used approach to probe cellular mechanisms. For example, to better understand the complex phenomenon of cancer drug resistance due to sequestration, binding and efflux, drug molecules are introduced into cancer cells. To probe cell-cell communication tracer molecules are delivered into connected cells and their junctional permeabilities are estimated. Existing methods for delivering molecules into cells include the injection of discrete boluses of material using mechanical or electro-mechanical principles, and continuous perfusion from a patch pipette in the whole cell configuration. However with these methods delivery cannot be quantified, or can introduce stress to the cells due to volume addition, or can cause excessive washout of cytosolic content. In this work we report on the use of a diffusional microburet that is a micropipette with a diffusion port at its tip to deliver molecules into an impaled target cell, quantifiably and with reduced washout of intracellular constituents. A new ratiometry technique called temporal ratiometry has also been developed in this work to provide dynamic time resolved concentration maps of the delivered molecules within the cell. These methods have been used to determine junctional permeabilities between connected A7r5 cells over a 30 min time frame. The DMB has also been used to dose the anticancer drug doxorubicin into the human breast cancer cell line MCF-7 and the rat epithelial cell line NRKE to titrate intracellular drug binding sites. Doxorubicin was found to accumulate in the nucleus and slow propagation at binding sites at the rate of ~2.0 ± 0.8 micrometers/min (n=4 cells) were observed for the first time. The methods presented in this work can make new types of experiment, such as continuous and quantitative titrations of binding sites, sequestration processes, and efflux pathways on single cells feasible. As an offshoot of diffusive dosing, we also report on the development of a delivery device that uses hydrodynamic resistance in the path of flow to effect very slow deliveries in the order of nL/s to make ultra low concentration solutions in a single step. Advisors/Committee Members: Gratzl, Miklos (Advisor).

Subjects/Keywords: Biomedical Research; Single cell; Diffusional drug delivery; Fluorescence; Ratiometry; Drug screening; Drug development; Cancer; Cancer drug resistance; Gap junctions; A7r5; NRKE; MCF7

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Oruganti, P. (2010). Drug Transport in Cell Preparations with Diffusional Dosing and Temporal Ratiometry. (Doctoral Dissertation). Case Western Reserve University School of Graduate Studies. Retrieved from http://rave.ohiolink.edu/etdc/view?acc_num=case1269536608

Chicago Manual of Style (16^th Edition):

Oruganti, Prasad. “Drug Transport in Cell Preparations with Diffusional Dosing and Temporal Ratiometry.” 2010. Doctoral Dissertation, Case Western Reserve University School of Graduate Studies. Accessed April 22, 2021. http://rave.ohiolink.edu/etdc/view?acc_num=case1269536608.

MLA Handbook (7^th Edition):

Oruganti, Prasad. “Drug Transport in Cell Preparations with Diffusional Dosing and Temporal Ratiometry.” 2010. Web. 22 Apr 2021.

Vancouver:

Oruganti P. Drug Transport in Cell Preparations with Diffusional Dosing and Temporal Ratiometry. [Internet] [Doctoral dissertation]. Case Western Reserve University School of Graduate Studies; 2010. [cited 2021 Apr 22]. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=case1269536608.

Council of Science Editors:

Oruganti P. Drug Transport in Cell Preparations with Diffusional Dosing and Temporal Ratiometry. [Doctoral Dissertation]. Case Western Reserve University School of Graduate Studies; 2010. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=case1269536608

4. Lin, Yu-Hung. Probing cellular mechano-sensitivity using biomembrane-mimicking cell substrates of adjustable stiffness.

Degree: 2015, IUPUI

URL: http://hdl.handle.net/1805/9965

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Indiana University-Purdue University Indianapolis (IUPUI)

It is increasingly recognized that mechanical properties of substrates play a pivotal role in the regulation of cellular fate and… (more)

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Indiana University-Purdue University Indianapolis (IUPUI)

It is increasingly recognized that mechanical properties of substrates play a pivotal role in the regulation of cellular fate and function. However, the underlying mechanisms of cellular mechanosensing still remain a topic of open debate. Traditionally, advancements in this field have been made using polymeric substrates of adjustable stiffness with immobilized linkers. While such substrates are well suited to examine cell adhesion and migration in an extracellular matrix environment, they are limited in their ability to replicate the rich dynamics found at cell-cell interfaces. To address this challenge, we recently introduced a linker-functionalized polymer-tethered multi-bilayer stack, in which substrate stiffness can be altered by the degree of bilayer stacking, thus allowing the analysis of cellular mechanosensitivity. Here, we apply this novel biomembrane-mimicking cell substrate design to explore the mechanosensitivity of C2C12 myoblasts in the presence of cell-cell-mimicking N-cadherin linkers. Experiments are presented, which demonstrate a relationship between the degree of bilayer stacking and mechanoresponse of plated cells, such as morphology, cytoskeletal organization, cellular traction forces, and migration speed. Furthermore, we illustrate the dynamic assembly of bilayer-bound N-cadherin linkers underneath cellular adherens junctions. In addition, properties of individual and clustered N-cadherins are examined in the polymer-tethered bilayer system in the absence of plated cells. Alternatively, substrate stiffness can be adjusted by the concentration of lipopolymers in a single polymer-tethered lipid bilayer. On the basis of this alternative cell substrate concept, we also discuss recent results on a linker-functionalized single polymer-tethered bilayer substrate with a lateral gradient in lipopolymer concentration (substrate viscoelasticity). Specifically, we show that the lipopolymer gradient has a notable impact on spreading, cytoskeletal organization, and motility of 3T3 fibroblasts. Two cases are discussed: 1. polymer-tethered bilayers with a sharp boundary between low and high lipopolymer concentration regions and 2. polymer-tethered bilayers with a gradual gradient in lipopolymer concentration.

Advisors/Committee Members: Naumann, Christoph A., Das, Chittaranjan, Thompson, David, Long, Eric C..

Subjects/Keywords: Artificial Substrate; Cadherin; Polymer-tethered Bilayer; Biomembrane-mimicking; Lipid Bilayer; Cadherins – Research; Cell junctions – Research; Polymers – Surfaces; Bilayer lipid membranes – Research; Cell adhesion – Research – Analysis; Biological interfaces – Research – Analysis; Cellular control mechanisms – Research; Cells – Mechanical properties – Research; Polymers – Rheology; Artificial cells – Research

…such as focal adhesions and adherens junctions.17-19 Consequently, polymeric cell substrates… …adherens junctions in polarized cells20,21. Herein, an alternative strategy for novel cell… …4.3.2.3 Cell Migration and Tortuosity… …23 Figure 2.2. 1 Cell migration on a 2D substrate… …29 Figure 2.2. 2 Complex of Integrin and surrounding proteins on cell membrane…

15 more images…

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Lin, Y. (2015). Probing cellular mechano-sensitivity using biomembrane-mimicking cell substrates of adjustable stiffness. (Thesis). IUPUI. Retrieved from http://hdl.handle.net/1805/9965

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Lin, Yu-Hung. “Probing cellular mechano-sensitivity using biomembrane-mimicking cell substrates of adjustable stiffness.” 2015. Thesis, IUPUI. Accessed April 22, 2021. http://hdl.handle.net/1805/9965.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Lin, Yu-Hung. “Probing cellular mechano-sensitivity using biomembrane-mimicking cell substrates of adjustable stiffness.” 2015. Web. 22 Apr 2021.

Vancouver:

Lin Y. Probing cellular mechano-sensitivity using biomembrane-mimicking cell substrates of adjustable stiffness. [Internet] [Thesis]. IUPUI; 2015. [cited 2021 Apr 22]. Available from: http://hdl.handle.net/1805/9965.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Lin Y. Probing cellular mechano-sensitivity using biomembrane-mimicking cell substrates of adjustable stiffness. [Thesis]. IUPUI; 2015. Available from: http://hdl.handle.net/1805/9965

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

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