高斯玻取样
Natural antisense transcripts(NATs)are widespread in mammalian genome, most of the NATs belong to noncoding RNA.In vivo,some of them have important biological functions, such as gene expression regulation, cell cycle regulation, cell differentiation, and so on. some NAT are relevant with tumor occurence. All-trans-retinoic acid (all-trans-retinoic acid, atRA) is a small molecule fat-soluble physical active material, which producted by vitamin A through two steps oxidation dehydrogenation (Retinol ↔ Retinal-aRA). Except for maintaining the normal development of embryos, system formation, atRA have a wide range of biological functions in maintaining adult animals growth, reproduce.And it can induce differentiation of malignant tumor cell, and change malignant tumor to the benign. NADP(H)-dependent retinol dehydrogenase/reductase (NRDR), encoded by DHRS4 gene, is a crucial enzyme in the synthesis of retinoic acid.Human DHRS4and other two homologous genes DHRS4L2、DHRS4L1,constitute a gene cluster located on the human chromosome 14q11.2. Genbank bioinformatics data shows that an antisense gene named C14orf167located at the 5 'end of DHRS4 gene, and they form a head to head overlapping structure. Our research group cloned an variant transc
步进式加热炉
ript of C14orf167, AS1DHRS4(1865bp),in HL7702 and liver cancer cell line HepG2, whose second exon overlap with the first exon of DHRS4. With HL7702, HepG2 cell lines as a model, they found that AS1DHRS4 regulated the DHRS4 gene clusters expression via epigenetic modifications. Expression of DHRS4is dysregulated in breast carcinoma, and expression of DHRS4and AS1DHRS4 present tissue special. However, whether AS1DHRS4 may play a role in regulation of DHRS4 gene expression in esophageal cancer cells, or whether DHRS4and AS1DHRS4 expression will be changed in esophageal carcinoma tissues are still unknown.
胎压监测装置To investigate the expression profile of AS1DHRS4 and its regulation role to DHRS4 gene in esophageal carcinorma, in the present study: (1) We cloned the antisense transcription, encoded by C14orf167 through 3'、5' RACE PCR in EC109 cells, which had the same sequences as AS1DHRS4；(2) AS1DHRS4 siRNA knocked down AS1DHRS4 in EC109 cells raised DHRS4 gene expression; (3) We constructed AS1DHRS4-pcDNA3.0 vector, and transfect it into the EC109 cells,overexpression of AS1DHRS4 did not change DHRS4 gene expression level;(4)Quantitative RT-polymerase chain reaction(QRT-PCR) showed that the transcription levels of AS1
金属化膜DHRS4 were higher than DHRS4in different esophageal carcinoma cell lines,tissues and paired non-cancer tissues. Compared with normal esophageal tissues, AS1DHRS4 transcription declined in esophageal carcinoma tissues.However, DHRS4gene expression had no distinct change；(5) We treated EC109, EC9706 cell lines with Trichostatin A (TSA) (a inhibitor of histone deacetylases), quantitative RT-PCR, Western blot results showed that TSA treatment resulted in increased AS1DHRS4 transcription, but decreased DHRS4 expression levels. Chromatin immunoprecipitation (CHIP) results indicated that TSA treatment decreased histone H3 acetylation in the promoter region of the DHRS4 gene, but increased H3 acetylation in the promoter region of the C14orf167 gene.
In summarry, we first identified an antisense transcript AS1DHRS4 existed in EC109 cells, and found that expression of AS1DHRS4 was much more than DHRS4both in esophageal cancer cell lines, tissues and paired normal esophageal tissues.AS1DHRS4 was decreased in esophageal cancer tissues. What is more, we demonstrated that histone modification regulates transcription of AS1DHRS4 in esophageal carcinoma cells. Histone acetylation induces AS1DHRS4 transcription to down-regulate DHRS4 expression.
Key words: DHRS4；AS1DHRS4；gene expression regulation
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