Investigation of caveolae-mediated endocytosis in response to interferon

interferon;virus;endocytosis;cholera toxin;caveolae;caveolin;cancer;flow cytometry;confocal microscopy

Yeager, Ashley ; Pack, Daniel W. ; Pack ; Daniel W. ; Kraft ; Mary L.

关键词: interferon;    virus;    endocytosis;    cholera toxin;    caveolae;    caveolin;    cancer;    flow cytometry;    confocal microscopy;   

Others  :  https://www.ideals.illinois.edu/bitstream/handle/2142/46906/Ashley_Yeager.pdf?sequence=1&isAllowed=y

美国|英语

来源: The Illinois Digital Environment for Access to Learning and Scholarship

【 摘 要 】

Cancer contributes substantially to overall mortality and morbidity worldwide, resulting in considerable research focused on treatments. Yet many types of cancers continue to have a poor prognosis [1]. Cancer is an incredibly diverse affliction, resulting from numerous changes in normal cell physiology. Treating such a diverse disease will take highly interdisciplinary work from clinicians and scientists to discover novel methods to fight this condition and improve existing treatments. The broad objective of this work is to obtain a better understanding of the actions of type I interferons (IFN), cytokines that display potent antiviral as well as anti-cancer activity. IFN has a variety of cellular effects, some of which lead to its anti-tumor nature, but many of which are not fully understood. Some cellular effects may contribute to the detrimental side effects that may occur during IFN treatment and significantly affect a patient’s quality of life. Thus, additional research is needed to improve the use of IFN as a cancer treatment.This work sheds light on type I interferon’s (IFN) effects on cellular endocytosis. Endocytosis, a vital process in healthy cells used to internalize material, is a likely IFN target. The anti-tumor nature of IFN may be an unintentional byproduct of its antiviral effects, which are designed to halt pathogenic assault, including pathogen entry into the cell via [2,3]. IFN-induced effects on endocytosis may also contribute substantially to its anti-tumor nature, as abnormal endocytosis has also been found in multiple cancers [4,5]. To examine the effects of IFN on endocytosis, several fluorescent techniques were used to quantify the internalization of cholera toxin in IFN-treated and untreated cells. Flow cytometry was used to quantify the internalization of fluorescent pathway-specific cholera toxin with establishedconcentrations [6], and the differences between the IFN-treated and untreated cell samples were assessed with statistical methods. The commercially available fluorescent reagent, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine, 4-chlorobenzenesulfonate (DiD) was also used to stain the plasma membrane and fluorescent-labeled cholera toxin to image IFN-treated and untreated cells with confocal microscopy. This work showed that IFN induced a statistically significant increase in the endocytosis of cholera toxin in NIH-3T3 cells.This work not only expands the basic knowledge of IFN and its cellular effects, but also further characterizes a potential IFN target, namely endocytosis. This knowledge may lead to its improved use as a cancer treatment, and may also benefit gene/drug delivery research. Elucidating these IFN-induced changes in endocytosis may also lead to improved therapies which mimic IFN’s effects while bypassing adverse side effects.