کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
3424190 | 1227199 | 2013 | 8 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Subcellular localization and quantitation of the human papillomavirus type 16 E6 oncoprotein through immunocytochemistry detection Subcellular localization and quantitation of the human papillomavirus type 16 E6 oncoprotein through immunocytochemistry detection](/preview/png/3424190.png)
Human papillomavirus (HPV) type 16 E6 is a viral oncoprotein essential for host cell transformation. Due to its role in HPV-induced cancers of the genital, head and neck epithelia, reliable protein-level determination of E6 expression would be an invaluable diagnostic tool. Immunocytochemical detection and subcellular localization of HPV16 E6 has been demonstrated with varying success and a comprehensive review of techniques is lacking. To address these issues, we used established monoclonal antibodies and optimized a standard immunocytochemical method for E6 protein detection inside the HPV16 positive cell lines, SiHa and CaSki. E6 oncoprotein was detected primarily in the nucleus. We also refined quantitative analysis with a software to objectively differentiate between HPV16 positive and negative cells. Our analysis was also able to differentiate expression differences between SiHa and CaSki on par with RT-qPCR. Thus, we provide a long-needed, robust protocol for antibody-mediated detection of the HPV16 E6 oncoprotein inside cultured cells.
► We report an immunocytochemical method for in situ HPV16 E6 protein detection.
► E6 protein is localized to the nucleus of carcinoma-derived cells.
► Reliable quantitative analysis of HPV16 E6 protein is achievable in cultured cells.
► Quantitative analysis is used to objectively measure protein-level E6 expression.
► Transcript and protein-level expression of E6 is consistent.
Journal: Virology - Volume 435, Issue 2, 20 January 2013, Pages 425–432