Quantitative real-time PCR analysis of YKL-40 and its comparison with mammalian Chitinase mRNAs in normal human tissues using a single standard DNA

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Abstract

© 2015 by the authors; licensee MDPI, Basel, Switzerland.YKL-40 (YKL for the first three N-terminal residues of a 40 kDa protein) belongs to a group of human chitinase-like proteins (CLPs), which are similar to chitinases but lack chitinolytic activity. YKL-40 mRNA and its protein levels have been reported elevated in multiple disorders including asthma, cystic fibrosis, rheumatoid arthritis and malignant tumors. Here, we quantified the YKL-40 mRNA levels and compared them with chitinases and housekeeping genes in normal human tissues. To establish the quantitative real-time PCR (qPCR) system for evaluation of relative YKL-40 mRNA levels, we constructed a human standard DNA molecule by ligating cDNAs of YKL-40, two mammalian chitinases and two housekeeping genes in a one-to-one ratio. We generated cDNAs from various normal human tissues and analyzed the YKL-40 mRNA expression levels using a qPCR system with the standard DNA. We found that YKL-40 mRNA is present widely in human tissues while its expression patterns exhibit clear tissue specificity. Highest YKL-40 mRNA levels were detected in the liver, followed by kidney, trachea and lung. The levels of YKL-40 mRNA in the kidney and liver were more than 100-times higher than those of chitotriosidase mRNA. Our study provides for the first time a comprehensive analysis of the relative expression levels of YKL-40 mRNA versus mammalian chitinases in normal human tissues.

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Ohno, M., Bauer, P. O., Kida, Y., Sakaguchi, M., Sugahara, Y., & Oyama, F. (2015). Quantitative real-time PCR analysis of YKL-40 and its comparison with mammalian Chitinase mRNAs in normal human tissues using a single standard DNA. International Journal of Molecular Sciences, 16(5), 9922–9935. https://doi.org/10.3390/ijms16059922

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