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Human dipeptidyl peptidase III mRNA variant I and II are expressed concurrently in multiple tumor derived cell lines and translated at comparable efficiency in vitro.

Mol. Biol. Rep.2016 Jun;43(6):457-62. Epub 2016 May 06
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摘要


Dipeptidyl peptidase III (DPP III) is an emerging biomarker of human cancers. Expression, specificity, and function of human DPP III (hDPP III) mRNA variant I (V-I), II (V-II), and III (V-III) are poorly understood. Here, we investigated expression of these variants in multiple human tumor derived cell lines. DNA sequencing revealed concurrent expression of hDPP III V-I and V-II in U87MG (glioblastoma), SCC4 (squamous cell carcinoma), SiHa (carcinoma of uterus) cells. In SKOV1 cells, a cell line derived from ovarian carcinoma where a positive correlation between histological aggressiveness of the malignancy and hDPP III expression has previously been established, only V-II could be detected. Human DPP III V-III, which lacks an in-frame coding sequence, could not be detected in any of these cell lines. 5' untranslated region (UTR) of hDPP III V-II contains nucleotides GCA (-12 to -10 bp) upstream to the translation initiator codon (AUG). These nucleotides are absent from V-I and V-III, however, both V-I and V-II encode for the same hDPP III protein isoform-I. In vitro transcription coupled translation assay using hDPP III V-I and V-II expression vectors which contained full length V-I and V-II cDNA including the variable cloned under T7 promoter, respectively revealed a comparable translational efficiency for both the variants, abrogating involvement of nucleotides GCA (-12 to -10 bp) in translation of the variants. Our results, for the first time, demonstrate concurrent expression in multiple tumor derived cell lines and a comparable in vitro translational efficiency for hDPP III V-I and II.

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