کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8533772 | 1560471 | 2018 | 24 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
An automated epifluorescence microscopy imaging assay for the identification of phospho-AKT level modulators in breast cancer cells
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کلمات کلیدی
EGFRTGF-βSPCASTIMsiNTTRPCpAktTRPVPMCAPFAPI3KPBSEGFHER2Phosphatidylinositol-4,5-bisphosphate 3-kinase - Phosphatidylinositol-4،5-bisphosphate 3-kinaseepithelial mesenchymal transition - انتقال مزانشیمی epithelialTransforming growth factor β - تبدیل فاکتور رشد βhigh content analysis - تجزیه و تحلیل محتوای بالاAssay development - تست توسعهEMT - تکنسین فوریتهای پزشکیGene silencing - خاموشی ژنBreast cancer - سرطان پستانepidermal growth factor - عامل رشد اپیدرمیPlasma membrane Ca2+-ATPase - غشای پلاسما Ca2 + -ATPasePhosphate buffered saline - فسفات بافر شورphosphatase and tensin homolog - فسفاتاز و تنسین همولوگphosphorylated Akt - فسفروئید آکتstromal interaction molecule - مولکول تعامل استروماAutomated microscopy - میکروسکوپ خودکارparaformaldehyde - پارافرمالدهیدtransient receptor potential vanilloid - پتانسیل ترانزیتی وینیلوئیدtransient receptor potential canonical - پتانسیل گیرنده گذرا کانونی استPten - ژن PTENCalcium channels - کانال های کلسیمHuman epidermal growth factor receptor 2 - گیرنده عامل فاکتور رشد اپیدرمی انسان 2Epidermal growth factor receptor - گیرنده فاکتور رشد اپیدرمال
موضوعات مرتبط
علوم پزشکی و سلامت
داروسازی، سم شناسی و علوم دارویی
داروشناسی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
AKT is an enzyme of the PI3K/pAKT pathway, regulating proliferation and cell survival. High basal levels of active, phosphorylated AKT (pAKT) are associated with tumor progression and therapeutic resistance in some breast cancer subtypes, including HER2 positive breast cancers. Various stimuli can increase pAKT levels and elevated basal pAKT levels are a feature of PTEN-deficient breast cancer cell lines. The aim of this study was to develop an assay able to identify modulators of pAKT levels using an automated epifluorescence microscope and high content analysis. To develop this assay, we used HCC-1569, a PTEN-deficient, HER2-overexpressing breast cancer cell line with elevated basal pAKT levels. HCC-1569 cells were treated with a selective pharmacological inhibitor of AKT (MK-2206) to reduce basal pAKT levels or EGF to increase pAKT levels. Immunofluorescence images were acquired using an automated epifluorescence microscope and integrated intensity of cytoplasmic pAKT staining was calculated using high content analysis software. Mean and median integrated cytoplasmic intensity were normalized using fold change and standard score to assess assay quality and to identify most robust data analysis. The highest zâ² factor was achieved for median data normalization using the standard score method (zâ²â¯=â¯0.45). Using our developed assay we identified the calcium homeostasis regulating proteins TPRV6, STIM1 and TRPC1 as modulators of pAKT levels in HCC-1569 cells. Calcium signaling controls a diverse array of cellular processes and some calcium homeostasis regulating proteins are involved in modulating pAKT levels in cancer cells. Thus, these identified hits present promising targets for further assessment.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Pharmacological and Toxicological Methods - Volume 92, JulyâAugust 2018, Pages 13-19
Journal: Journal of Pharmacological and Toxicological Methods - Volume 92, JulyâAugust 2018, Pages 13-19
نویسندگان
Elke Kaemmerer, Dane Turner, Amelia A. Peters, Sarah J. Roberts-Thomson, Gregory R. Monteith,