A quantitative study on splice variants of N-acylethanolamine acid amidase in human prostate cancer cells and other cells

Biochim Biophys Acta. 2016 Dec;1861(12 Pt A):1951-1958. doi: 10.1016/j.bbalip.2016.09.018. Epub 2016 Oct 5.

Abstract

N-Acylethanolamine acid amidase (NAAA) is a lysosomal enzyme, hydrolyzing various bioactive N-acylethanolamines with a preference for palmitoylethanolamide. Human NAAA mRNA was previously reported to consist of multiple 3'-end splice variants. However, their tissue distributions and roles have not been examined yet. In the present study, we first identified four major splice variants (tentatively referred to as a1, a2, b2, and c2) in a human prostate cancer cell line LNCaP, which were composed of exons 1-11, exons 1-10 and 12, exons 1-9 and 12, and exons 1-8 and 12, respectively. We next developed quantitative polymerase chain reaction methods to individually quantify these NAAA variants as well as collectively measure all the variants. Among various human prostate cancer cells, the total levels of NAAA mRNAs in androgen-sensitive cells like LNCaP were higher than those in androgen-insensitive cells. In all of these prostate cells and other human cells, variants a1 and b2 showed the highest and lowest expression levels, respectively, among the four variants. Interestingly, ratios of the four variants were different by cell type. Variants a1 and a2 encoded the same full-length NAAA protein, which was catalytically active, while b2 and c2 were translated to C-terminally truncated proteins. As expressed in HEK293 cells these truncated forms were detected as catalytically inactive precursor proteins, but not as mature forms. These results revealed wide distribution of multiple variants of NAAA mRNA in various human cells and suggested that the proteins from some variants are catalytically inactive.

Keywords: Endocannabinoid; N-Acylethanolamine acid amidase; Prostate cancer; Quantitative PCR; Splice variant.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidohydrolases / genetics*
  • Ethanolamines / metabolism
  • Evaluation Studies as Topic
  • Exons / genetics
  • Genetic Variation / genetics*
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • MCF-7 Cells
  • Male
  • Prostatic Neoplasms / genetics*
  • Prostatic Neoplasms / metabolism
  • RNA Splicing / genetics*
  • RNA, Messenger / genetics

Substances

  • Ethanolamines
  • N-acylethanolamines
  • RNA, Messenger
  • Amidohydrolases
  • NAAA protein, human