Detection of CK19 Marker in the MCF7 and T47D Cell Lines by Immunofluorescence Microscopy

Publish Year: 1393
نوع سند: مقاله کنفرانسی
زبان: English
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ICBCMED10_057

تاریخ نمایه سازی: 21 اردیبهشت 1397

Abstract:

Cytokeratin19 (CK19) is a part of the cytoskeleton of epithelial cells. It is a sensitive and specific marker of epithelial cells that expressed in breast cancer cells. Immunofluorescence microscopy is a powerful technique that is widely used to assess both the localization and expression levels of proteins. On the other hand, T47D and MCF7 are two human hormone-dependent breast cancer cell lines, which are widely used as experimental models for breast cancer studies. Materials and methods: breast cancer cell lines (MCF7, T47D) were cultured in RPMI 1640 containing 10% fetal bovine serum (FBS) and penicillin-streptomycin antibiotic. Then, cells were fixed with 4% paraformaldehyde and permeabilized with ice-cold 100% methanol. The cells were blocked with 1% BSA and were stained with CK19 antibody. Finally, CK19 expression was elevated by mmunofluorescence microscopy.Results: Immunofluorescence microscopy was used to detect the CK19 expression in T47D and MCF7 cells. The result showed that T47D and MCF7 cells were CK19-immunoreactive cells and CK19 was mainly located in the cytoplasm of these cells.Conclusion: T47D and MCF7 cell lines belonged to luminal A subtype of breast cancer that CK19 expression correlated whit ER (Estrogen) marker. Expression of CK19 in the estrogen positive (ER+) breast cancer cell lines suggest that these cells have originated from luminal epithelial cells.CK19 is an epithelial marker that estrogen regulated its expression in luminal A subtype of breast cancer cell lines.

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Authors

Zahra Orafa

Master of Science, Department of Molecular and Cellular Sciences, Pharmaceutical Sciences Branch,Islamic Azad University, Tehran, Iran

Mana Oloomi

Associate Professor, Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran

Saeideh Keyvani

Master of Science, Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran

Nasrin Karimi

Master of Science, Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran