OBJECTIVETo investigate the possible biological function and mechanism of miR-143 in the metastasis of human nasopharyngeal carcinoma (NPC).

METHODSUsing bioinformatics to predict the target gene of miR-143, the 3'UTR and mutant 3'UTR of GLI3 gene was cloned into psiCHECK-2 vector. Dual-luciferase reporter gene assay was employed to examine the repression of the GLI3 gene. miR-143 and GLI3 expression levels in 5-8F cells transfected with miR-143 mimics, inhibitor, or siGLI3 were examined, and the changes in the cell migration ability was assessed by Transwell invasion assay.

RESULTSBioinformatics prediction indicated the Hh pathway transcription gene GLI3 as a target gene of miR-143, and dual-luciferase reporter assay showed that miR-143 directly combined with the 3'UTR of GLI3. qRT-PCR and Western blotting demonstrated that the expression of miR-143 in 5-8F cells was negatively correlated to GLI3 and suppressed the migration of 5-8F cells.

CONCLUSIONMiR-143 can inhibit the invasion of NPC cells by negative regulation of GLI3 gene, which sheds light on the role of miR-143 and Hh pathway in NPC.

Carcinoma ; Cell Line, Tumor ; Cell Movement ; genetics ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Genes, Reporter ; Humans ; Kruppel-Like Transcription Factors ; genetics ; MicroRNAs ; genetics ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Nerve Tissue Proteins ; genetics ; Zinc Finger Protein Gli3

Objective:To investigate the characteristics of urinary microflora in women with type 2 diabetic peripheral neuropathy without lower urinary tract symptoms.Methods:By completing nerve conduction function and the American Urological Association Symptom Index questionnaire (AUA-SI), a total of 30 cases of women hospitalized with type 2 diabetes and no symptoms of lower urinary tract from May 2017 to August 2018 were included. 17 patients with diabetic peripheral neuropathy were assigned to the DPN group, and 13 patients without diabetic peripheral neuropathy were assigned to the nDPN group. Urine specimens were collected from clean catch midstream urine and processed for extracting DNA. Microbial diversity and composition were analyzed using the Illumina sequencing platform targeting to 16S rDNA gene. Sequencing reads were processed by QIIME. LEfSe algorithm was used to analyze the flora with significant differences between the two groups.Results:The duration of diabetes in the DPN group was lower than that in the nDPN group [(4.12 ± 3.28)years vs.(8.03 ± 6.11)years, P = 0.03], and the retinopathy cases were more in the DPN group than those in the nDPN group (6 vs. 0, P=0.03). Except for above two indicators, there was no significant difference in demographic characteristics between DPN group and nDPN group( P>0.05). The urinary microenvironment of DPN was characterized by increased bacterial richness(sobs index, chao index and aec index, 67.24±40.25 vs.108.69±57.18; 81.36±47.99 vs.122.55±55.70; 88.58±55.03 vs.125.78±53.03, all P<0.05) and by the enrichment of Mycoplasmataceae(Metastats value: 0.52±0.01vs.0.01±0.00001, P=0.02). Beta diversity showed that no significant difference of bacterial composition was found between these two group( P>0.05). LEfSe analysis showed that at the genus level, the relative abundance of eight genera(e.g., Bacillus, Duganella, Leptotrichia, Proteus, Propionibacterium, Pseudoxanthomonas, Bdellovibrio and uncultured_soil_bacterium) in DPN group decreased at the level of genus( P<0.05). Conclusions:Female patients with type 2 diabetes without lower urinary tract symptoms of peripheral neuropathy exhibit a different microbial community compared to nDPN controls. Mycoplasmataceae may be a potential biomarker for patients with DPN.