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Tumor Biology

, Volume 37, Issue 8, pp 11115–11126

First Online: 24 February 2016Received: 29 October 2015Accepted: 11 February 2016

Abstract

In this study, we analyzed the expression profile of four genes CCNA2, CCNB1, CCNB2, and CDK1 in laryngeal squamous cell carcinoma LSCC cell lines and tumor samples. With the application of microarray platform, we have shown the overexpression of these genes in all analyzed LSCC samples in comparison to non-cancer controls from head and neck region. We have selected CDK1 for further analysis, due to its leading role in cell cycle regulation. It is a member of the Ser-Thr protein kinase family of proven oncogenic properties. The results obtained for CDK1 were further confirmed with the application of reverse transcription quantitative polymerase chain reaction RT-qPCR technique, Western blot, and immunohistochemistry IHC. The observed upregulation of CDK1 in laryngeal squamous cell carcinoma has encouraged us to analyze for genetic mechanisms that can be responsible this phenomenon. Therefore, with the application of array-CGH, sequencing analysis and two methods for epigenetic regulation analysis DNA methylation and miRNA expression, we tried to identify such potential mechanisms. Our attempts to identify the molecular mechanisms responsible for observed changes failed as we did not observe significant alterations neither in the DNA sequence nor in the gene copy number that could underline CDK1 upregulation. Similarly, the pyrosequencing and miRNA expression analyses did not reveal any differences in methylation level and miRNA expression, respectively; thus, these mechanisms probably do not contribute to elevation of CDK1 expression in LSCC. However, our results suggest that alteration of CDK1 expression on both mRNA and protein level probably appears on the very early step of carcinogenesis.

KeywordsCDK1 Overexpression Gene methylation microRNA Western blot Immunohistochemistry Electronic supplementary materialThe online version of this article doi:10.1007-s13277-016-4991-4 contains supplementary material, which is available to authorized users.

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Author: K. Bednarek - K. Kiwerska - M. Szaumkessel - M. Bodnar - M. Kostrzewska-Poczekaj - A. Marszalek - J. Janiszewska - A. Bart

Source: https://link.springer.com/article/10.1007/s13277-016-4991-4







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