Human salivary cystatin S. Cloning, sequence analysis, hybridization in situ and immunocytochemistry.
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ABSTRACT:
SUBMITTER: Bobek LA
PROVIDER: S-EPMC1151393 | biostudies-other | 1991 Sep
REPOSITORIES: biostudies-other
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Project description:Objective: Salivary rare basaloid lesions, including cribriform type basal cell adenoma (cBCA), BCA with incomplete capsule (iBCA), sialoblastoma (SB), and intercalated duct hyperplasia (IDH), could easily be misdiagnosed as adenoid cystic carcinoma (AdCC). We aim to identify an approach for differential diagnosis and to establish an optimal workflow concerning the diagnosis of these lesions. Material and methods: A panel of antibodies (MYB, ?-catenin, CD117, SOX10, ki67, P63, calponin) and fluorescence in situ hybridization (FISH)-MYB were utilized to distinguish above salivary basaloid diseases from AdCC. Results: Histologically, the striking diagnostic features of cBCA, iBCA, SB, and IDH are composed of basaloid tumor cells, well-defined encapsulation, or lack of destructive invasion. Immunohistochemically, Myb immune-labeling could effectively make a distinction among cBCA, iBCA, SB, and IDH from AdCC, except in SB. cBCA and iBCA typically expressed ?-catenin in the nuclei of tumor cells. There was no statistical significance in the ki67 index between SB and AdCC, but their indices were significantly higher than those of iBCA and IDH (p < 0.05, p < 0.05, respectively). P63 and calponin immune-expression were observed in the basaloid or myoepithelial cells. CD117 were observed positively in cBCA, iBCA, SB, and AdCC, except in IDH. SOX10 were observed positively in all cases. No cases had fusion of MYB and NFIB detectable by FISH, except in AdCC. Conclusion: Considering their sensitivity and specificity, FISH-Myb and an immunohistochemical panel of MYB/?-catenin/ki67 would be an optimal choice for the differential diagnosis of these basaloid lesions. Clinical relevance: Some salivary basaloid tumor or tumor-like lesions have overlapping features with AdCC. Through this present research, we suggested that the panel IHC of MYB, ?catenin, and ki67 combined with FISH-Myb should be an optimal choice for differential diagnosis among those lesions.
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Project description:ObjectiveTo amplify the entire ORF of SLC25A13 cDNA which encodes citrin, a liver-type mitochondrial aspartate-glutamate carrier, and to investigate sequence feature of the transcripts for this gene in cultured human amniocytes. This study will provide laboratory evidences for prenatal diagnosis of NICCD at mRNA level.MethodsTotal RNA was extracted from cultured amniocytes, and cDNA was synthesized, and then nest PCR was performed to amplify the entire ORF sequences of SLC25A13. The PCR products were purified, cloned, sequenced, and aligned with the genomic DNA of SLC25A13 to analyze the alternative splicing pattern.ResultsThe entire ORF of SLC25A13 gene was successfully amplified. Three splice variants of SLC25A13, i.e., SLCA (normal mRNA), SLCB (CAG insertion between exon 9-10) and SLCC (exon 5-11 skipping), were identified in the subjects. However, no abnormal mRNA from the allele with mutation 851del4 was detected in the amniocytes cultured from a carrier fetus of this mutation.ConclusionsThis study demonstrated that the entire ORF of SLC25A13 cDNA can be successfully amplified from cultured human amniocytes, and revealed exon 5-11 skipping as a novel SLC25A13 transcript. Normal mRNA occupied majority of the transcripts in normal control and heterozygous amniocytes which contained normal allele and 851del4 mutation, indicating that the two fetuses wouldn't suffer from NICCD. These SLC25A13 transcription features provided laboratory evidences for prenatal diagnosis of NICCD.
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