OBJECTIVE: To explore the genetic basis for a Chinese patient featuring cleidocranial dysplasia(CCD). METHODS: Genomic DNA was extracted from peripheral blood samples of the patient and his parents. Whole exome sequencing (WES) was carried out for the patient, and suspected variant was verified by Sanger sequencing. RESULTS: WES has identified a missense c.460G>T (p.Val154Phe) (GRCh37/hg19) variant of the RUNX2 gene. The variant was located in the Runt domain, a highly conserved region (PM1); it was not present in either the Genome Aggregation Database or the 1000 Genomes Project (PM2), and was predicted to have a deleterious effect on the gene product by multiple in silico prediction tools (PP3); the clinical phenotype of the patient was highly consistent with that of cleidocranial dysplasia (PP4). Furthermore, the variant was unreported in medical literature and was absent in both parents (PS2). Based on the American College of Medical Genetics and Genomics guidelines, the c.460 G>T variant of RUNX2 gene was predicted to be pathogenic (PS2+PM1+PM2+PP3+PP4). CONCLUSION The c.460G>T (p.Val154Phe) variant of the RUNX2 gene probably underlay the clinical phenotype in the patient. Above finding has enabled accurate diagnosis and expanded the spectrum of RUNX2 variants.
China ; Cleidocranial Dysplasia/genetics* ; Core Binding Factor Alpha 1 Subunit/genetics* ; Humans ; Mutation ; Whole Exome Sequencing

Objective To observe the effects of diacetyldianhydrogalactito (DADAG) on the proliferation and apopotosis in lung cancer cell NCI-H460.Methods MTT assay was performed to determine the half inhibitory concentration of DADAG(2.88,5.75,11.50,23.00 and 46.00 μg/mL)for the NCI-H460 cells and colony formation assay was used to detect the ability of cell proliferation; through AO/EB staining method, morphological changes of apoptotic cell under fluorescent microscope were observed;cell apoptosis was detected by flow cytometry to test the effects of DADAG on NCI-H460;RT-PCR was used to detect the effect of DADAG on Bcl-2 and Bax mRNA expres-sion levels within the cell. Results Compared with that in blank control group, cell proliferation was inhibited in the group treated with DADAG; the number of colony formation decreased and AO/EB staining results showed that cell apoptosis was characterized by typical morphological changes such as swelling, shrinking and fragmentation. Flow cytometry detection results showed that apoptosis rate was increased in the group treated with DADAG(all P<0.05).RT-PCR results indicated that the expression of pro-apoptosis gene Bax was up-regulated,while expression of anti-apoptosis gene Bcl-2 down-regulated. Conclusions DADAG exhibits the inhibitory activity in lung cancer cell NCI-H460 and can induce the apoptosis.The potential mechanism maybe associated with up-regulating the ex-pression of Bax but down-regulating the expression of Bcl-2.